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2007

Guide to

Biotechnology
http://www.pacontrol.com/introduction-to-biotechnology.htmlhttp://www.pacontrol.com/introduction-to-biotechnology.htmlhttp://www.pacontrol.com/introduction-to-biotechnology.htmlhttp://www.pacontrol.com/introduction-to-biotechnology.html

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The Guide to Biotechnologyis compiled by the Biotechnology Industry

Organization (BIO)

Debbie Strickland, BIO, Director of Marketing, Editor

Contributors

Deb Carstoiu, BIO, Director of State Media Relations and Advocacy

Elinor Van Dyck, Blue House Publishing, Art Director

Barbara Glenn, BIO, Managing Director of Animal Bitoechnology

Crispin Littlehales, Writer/Editor

Adrienne Massey, Ph.D., Writer/Editor

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Guide to Biotechnology n Biotechnology Industry Organization

Contents

Biotechnology: A Collection

of Technologies 1

What Is Biotechnology? ......................................................1Cells and Biological Molecules ...........................................1

Biotechnology Industry Facts 2

Market Capitalization, 19942005 .....................................3

U.S. Industry Statistics: 19942005 ...................................3

New Biotech Drug and Vaccine Approvals/New Indication Approvals by Year.................................4

North American Biotech Companiesby State and Province ....................................................4

Total Financing, 19982005 ...............................................5

Biotech Industry Financing, 2005 .....................................5

Time Line 6

Biotechnology Policy Milestones .....................................16

The Technologies

and Their Applications 18

Bioprocessing Technology ................................................18

Monoclonal Antibodies .....................................................18

Cell Culture ......................................................................19

Recombinant DNA Technology ........................................21

Cloning .............................................................................21

Protein Engineering ........................................................22

Biosensors ........................................................................22

Nanobiotechnology...........................................................23

Microarrays .......................................................................23

Biotechnology Tools in Research

and Deelopment 25

Research Applications ......................................................25Includes stem cell technology, cloning, microarraytechnology, antisense and RNA intererence, and gene

knockouts

Putting the Pieces Together: Omics ...............................29Genomics, proteomics and bioinormatics

Product Development Applications .................................31

Health-Care Applications 34

Diagnostics .......................................................................34

Therapeutics .....................................................................34

Personalized Medicine ......................................................37

Regenerative Medicine .....................................................39

Vaccines ............................................................................39

Plant-Made Pharmaceuticals ............................................41

Therapeutic Deelopement Oeriew 42

Biotech Drug Development Process ................................44

Approed Biotechnology Drugs 45

Agricultural Production Applications 65

Crop Biotechnology ..........................................................65Includes production improvement, impact on develop-ing countries, environmental and economic benets,

and regulations

Forest Biotechnology .......................................................69

Animal Biotechnology ......................................................70Includes companion animals, applications or humanmedicine, and environmental and conservation eorts

Aquaculture ......................................................................76

Global Area o Transgenic Crops,1995 to 2005: Industrial and Developing Countries ..78

Global Area o Transgenic Crops in 2004 and 2005by Country ..................................................................78

Global Area o Transgenic Crops in 2004 and2005 by Crop ...............................................................79

Global Area o Transgenic Crops, 1996 to 2005,by Crop ........................................................................79

Global Area o Transgenic Crops in 2004 and2005 by Trait ...............................................................80

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Biotechnology Industry Organization nGuide to Biotechnology

Global Area o Transgenic Crops, 1995 to 2005,by Trait ........................................................................80

Transgenic Crop Area as Percentage oGlobal Area o Principal Crops ....................................81

Global Status o Biotech Crops in 2005 ...........................81

Agricultural Biotech Products

on the Market 82

Food Biotechnology 87

Improving the Raw Materials ...........................................87

Food Processing................................................................88

Food Saety Testing ..........................................................89

Industrial and Enironmental

Applications 90

Industrial Sustainability ................................................... 90

Biocatalysts .......................................................................91

Biouel ..............................................................................92

Green Plastics ...................................................................93

Nanotechnology ................................................................93

Environmental Biotechnology .........................................94

Industries That Benet.....................................................95

Some Industrial Biotech Applicationsby Sector ......................................................................95

Consumer Products Made

with Industrial Biotechnology 96

Examples of Industrial Enzymes 98

Preparedness for

Pandemics and Biodefense 100

Policy...............................................................................100

A Strategic Asset .............................................................100Vaccines, monoclonal antibodies, DNA- or RNA-basedtherapeutics, and detection and diagnosis

Other Approaches ...........................................................101

Other Uses 102

DNA Fingerprinting........................................................102

Ethics 104

BIO Activities ..................................................................104

Ethical Issues ..................................................................105

BIO Statement of Ethical Principles 109

Intellectual Property 111

What Is a Patent? ............................................................111

The Purpose o a Patent .................................................111Patentable Inventions .....................................................112

Patent Requirements ......................................................112

The Patent Application ...................................................113

Patenting Organisms ......................................................113

Patent Licensing .............................................................114

Biotechnology Resources 115

Periodicals, Headline Services and Web Sites ...............115

General Science Journals ..............................................116

Biotech Education & Careers .........................................116

Selected Recent Reports on Biotechnology ...................116

Glossary 119

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Biotechnology: A Collection o Technologies

What Is Biotechnology?

Break biotechnology into its rootsand you have

biothe use of biological processes; and

technologyto solve problems or make useful products.

Using biological processes is hardly a noteworthyevent. We began growing crops and raising animals10,000 years ago to provide a stable supply o oodand clothing. We have used the biological processes omicroorganisms or 6,000 years to make useul oodproducts, such as bread and cheese, and to preservedairy products. Why is biotechnology suddenly receiv-ing so much attention?

During the 1960s and 70s our understanding o biol-ogy reached a point where we could begin to use thesmallest parts o organismstheir biological mol-eculesin addition to using whole organisms.

A more appropriate denition in the new sense o theword is this:

New Biotechnologythe use of cellular andbiomolecular processes to solve problems or makeuseful products.

We can get a better handle on the meaning o the wordbiotechnology by simply changing the singular nounto its plural orm, biotechnologies.

Biotechnology is a collection o technologies that capi-talize on the attributes o cells, such as their manuac-turing capabilities, and put biological molecules, suchas DNA and proteins, to work or us.

Cells and Biological Molecules

Cells are the basic building blocks o all living things.

The simplest living things, such as yeast, consisto a single, sel-sucient cell. Complex creatures moreamiliar to us, such as plants, animals and humans, aremade o many dierent cell types, each o which perormsa very specic task.

In spite o the extraordinary diversity o cell types inliving things, what is most striking is their remarkablesimilarity. This unity o lie at the cellular level providesthe oundation or biotechnology.

All cells have the same basic design, are made o the sameconstruction materials and operate using essentially thesame processes. DNA (deoxyribonucleic acid), the genetic

material o almost all living things, directs cell construc-tion and operation, while proteins do all the work. Be-cause DNA contains the inormation or making proteins,it directs cell processes by determining which proteinsare produced and when.

All cells speak the same genetic language. The DNA inor-mation manual o one cell can be read and implementedby cells rom other living things. Because a geneticinstruction to make a certain protein is understood bymany dierent types o cells, technologies based on cellsand biological molecules give us great fexibility in usingnatures diversity.

In addition, cells and biological molecules are extraordi-narily specic in their interactions. As a result, biotech-nology products can oten solve specic problems, gener-ate gentler or ewer side eects and have ewer unintendedconsequences.Specic, precise, predictable. Those are the

words that best describe todays biotechnology.

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2


n The biotechnology industry originated in the 1970s,based largely on a new recombinant DNA technique

whose details were published in 1973 by Stanley

Cohen o Stanord University and Herbert Boyer o theUniversity o Caliornia, San Francisco. RecombinantDNA is a method o making proteinssuch as humaninsulin and other therapiesin cultured cells undercontrolled manuacturing conditions. Boyer went onto co-ound Genentech, which today is biotechnologyslargest company by market capitalization.

n Biotechnology has created more than 200 new thera-pies and vaccines, including products to treat cancer,diabetes, HIV/AIDS and autoimmune disorders.

n There are more than 400 biotech drug products and

vaccines currently in clinical trials targeting morethan 200 diseases, including various cancers, Alzheim-ers disease, heart disease, diabetes, multiple sclerosis,

AIDS and arthritis.

n Biotechnology is responsible or hundreds omedi-cal diagnostic tests that keep the blood supply saerom the AIDS virus and detect other conditions earlyenough to be successully treated. Home pregnancytests are also biotechnology diagnostic products.

n Consumers are enjoying biotechnology oods such aspapaya, soybeans and corn. Biopesticides and other

agricultural products also are being used to improveour ood supply and to reduce our dependence onconventional chemical pesticides.

n Environmental biotechnology products make it pos-sible to clean up hazardous waste more eciently byharnessing pollution-eating microbes without the useo caustic chemicals.

n Industrial biotechnology applications have led to cleanerprocesses that produce less waste and use less energy and

water in such industrial sectors as chemicals, pulp andpaper, textiles, ood, energy, and metals and minerals. For

example, most laundry detergents produced in the UnitedStates contain biotechnology-based enzymes.

n DNA ngerprinting, a biotech process, has dramati-cally improved criminal investigation and orensicmedicine, as well as aorded signicant advances inanthropology and wildlie management.

n The biotech industry is regulated by the U.S. Food andDrug Administration (FDA), the Environmental Pro-tection Agency (EPA) and the Department o Agricul-ture (USDA).

n As o Dec. 31, 2005, there were 1,415 biotechnologycompanies in the United States, o which 329 werepublicly held.

n Market capitalization, the total value o publicly tradedbiotech companies (U.S.) at market prices, was $410billion as o Dec. 31, 2005.

n The biotechnology industry has mushroomed since1992, with U.S. health-care biotech revenues increas-ing rom $8 billion in 1992 to $50.7 billion in 2005.

n Biotechnology is one o the most research-intensiveindustries in the world. The U.S. biotech industryspent $19.8 billion on research and development in2005.

n The top ve biotech companies invested an average o$130,000 per employee in R&D in 2005.

n In 1982, recombinant human insulin became the rstbiotech therapy to earn FDA approval. The product

was developed by Genentech and Eli Lilly and Co.

n Corporate partnering has been critical to biotechsuccess. In 2005, biotech companies signed 564 newagreements with pharmaceutical rms and 354 withellow biotechs, according to BioWorld.

n Most biotechnology companies are young companies

developing their rst products and depend on

investorcapital or survival. Biotechnology attracted more than$20 billion in nancing in 2005 and has raised morethan $100 billion since 2000.

n The biosciencesincluding not just biotechnologybut all lie sciences activitiesemployed 1.2 millionpeople in the United States in 2004 and generated anadditional 5.8 million related jobs.

n The average annual wage o U.S. bioscience workerswas $65,775 in 2004, more than $26,000 greater thanthe average private sector annual wage.

n Bioethanolmade rom crop wastes using biotechenzymescould meet a quarter o U.S. energy needsby 2025.

n The Biotechnology Industry Organization (BIO) wasounded in 1993 to represent biotechnology com-panies at the local, state, ederal and internationallevels. As o December 2006, BIOs membership con-sisted o more than 1,100 biotechnology companies,academic centers, state and local associations andrelated enterprises.

Biotechnology Industry Facts

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Market Capitalization, 19942005*

*Amounts are U.S. dollars in billions.

Sources:Ernst & Young LLP

BioWorld

Year 2005 2004 2003 2002 2001 2000 1999 1998 1997 1996 1995 1994

Sales 32.1 28.1 28.4 24.3 21.4 19.3 16.1 14.5 13 10.8 9.3 7.7

Reenues 50.7 43.8 39.2 29.6 29.6 26.7 22.3 20.2 17.4 14.6 12.7 11.2

R&D Expense 19.8 19.6 17.9 20.5 15.7 14.2 10.7 10.6 9.0 7.9 7.7 7.0

Net Loss 4.1 6.8 5.4 9.4 4.6 5.6 4.4 4.1 4.5 4.6 4.1 3.6

No. of Public

Companies329 331 314 318 342 339 300 316 317 294 260 265

No. of Companies 1,415 1,346 1,473 1,466 1,457 1,379 1,273 1,311 1,274 1,287 1,308 1,311

U.S. Biotech Industry Statistics: 19942005*

*Amounts are U.S. dollars in billions.

Source:

Ernst & Young LLP, annual biotechnology industry reports, 19952006. Financial data based primarily on scal-year nancial statements of publiclytraded companies.

1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005

0

50

100

150

200

250

300

350

400

450

Year

45 4152

8393

137.9

353.5

330.8

225206

336.8

410

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4


New Biotech Drug and Vaccine Approvals/New Indication Approvals by Year

Source:

BIO

1982 1983 1984 1985 1986 1987 1988 1989 1990 1991 1992 1993 1994 1995 1996 1997 1998 1999 2000 2001 2002 2003 2004 2005

0

5

10

15

20

25

30

35

40

NumberofApprovals

Year

2

0 01

5

3 3

65

9

4

7 7

15

25

19

25

20

34

25

3637

40

38

California

Massachusetts

Ontario

Quebec

NorthCarolina

Maryland

BritishColumbia

NewJersey

Pennsylvania

Georgia

NewYork

Texas

Florida

Alberta

Washington

Connecticut

0

50

100

150

200

250

300

350

400

NumberofCompanies

Regions

374

256

143134

81 75 74 6859 58

56 48 47 45 33 25

North American Biotech Companies by State and Province

Source:

Ernst & Young LLP

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1998 1999 2000 2001 2002 2003 2004 2005

0

5

10

15

20

25

30

35

40

5.4

11.8

38

15.1

10.5

16.9

20.8 20.1

Total Financing, 19982005 (in billions of U.S. dollars)

Source:

BioWorld

Biotech Industry Financing, 2005

Total: $20,114.9 Million(all gures in millions)

Source:

BioWorld

Other nancings ofpublic companies:

$9,726.4(48.4%)

Public offerings:

$5,579.6

(27.7%)

Venture funding:

$4,808.9(23.9%)

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6


8000 B.C.

n Humans domesticate crops and livestock.

n

Potatoes rst cultivated or ood.

40002000 B.C.n Biotechnology rst used to leaven bread and erment

beer, using yeast (Egypt).

n Production o cheese and ermentation o wine (Sume-ria, China and Egypt).

n Babylonians control date palm breeding by selectivelypollinating emale trees with pollen rom certain maletrees.

500 B.C.n First antibiotic: moldy soybean curds used to treat

boils (China).

A.D. 100

n First insecticide: powdered chrysanthemums (China).

1322

n An Arab chietain rst uses articial insemination toproduce superior horses.

1590n Janssen invents the microscope.

1663

n Hooke discovers existence o the cell.

1675

n Leeuwenhoek discovers bacteria.

1761

n Koelreuter reports successul crossbreeding o cropplants in dierent species.

1797

n Jenner inoculates a child with a viral vaccine toprotect him rom smallpox.

18301833

n 1830Proteins discovered.

n 1833First enzyme discovered and isolated.

18351855

n Schleiden and Schwann propose that all organisms arecomposed o cells, and Virchow declares, Every cell

arises rom a cell.

1857n Pasteur proposes microbes cause ermentation.

1859n Charles Darwin publishes the theory o evolution by

natural selection. The concept o careully select-ing parents and culling the variable progeny greatlyinfuences plant and animal breeders in the late 1800sdespite their ignorance o genetics.

1865

n Science o genetics begins: Austrian monk GregorMendel studies garden peas and discovers that genetictraits are passed rom parents to ospring in a predict-able waythe laws o heredity.

18701890

n Using Darwins theory, plant breeders crossbreed cot-ton, developing hundreds o varieties with superiorqualities.

n Farmers rst inoculate elds with nitrogen-xing

bacteria to improve yields.

n William James Beal produces rst experimental cornhybrid in the laboratory.

n 1877A technique or staining and identiying bacte-ria is developed by Koch.

n 1878The rst centriuge is developed by Laval.

n 1879Fleming discovers chromatin, the rod-likestructures inside the cell nucleus that later came to becalled chromosomes.

1900

n Drosophila (ruit fies) used in early studies o genes.

1902n The term immunology rst appears.

1906n The termgenetics is introduced.

Time Line

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1911

n The rst cancer-causing virus is discovered by Rous.

1914

n Bacteria are used to treat sewage or the rst time inManchester, England.

1915

n

Phages, or bacterial viruses, are discovered.

1919

n First use o the word biotechnology in print.

1920

n The human growth hormone is discovered by Evansand Long.

1928

n Penicillin discovered as an antibiotic: Alexander Fleming.

n A small-scale test o ormulatedBacillus thuringiensis(Bt) or corn borer control begins in Europe. Commercialproduction o this biopesticide begins in France in 1938.

n Karpechenko crosses radishes and cabbages, creatingertile ospring between plants in dierent genera.

n Laibach rst uses embryo rescue to obtain hybridsrom wide crosses in crop plantsknown today ashybridization.

1930

n U.S. Congress passes the Plant Patent Act, enablingthe products o plant breeding to be patented.

1933

n Hybrid corn, developed by Henry Wallace in the 1920s, iscommercialized. Growing hybrid corn eliminates the op-tion o saving seeds. The remarkable yields outweigh theincreased costs o annual seed purchases, and by 1945,hybrid corn accounts or 78 percent o U.S.-grown corn.

1938n The termmolecular biology is coined.

1941n The termgenetic engineering is rst used, by Danish

microbiologist A. Jost in a lecture on reproduction inyeast at the technical institute in Lwow, Poland.

1942

n The electron microscope is used to identiy and char-acterize a bacteriophagea virus that inects bacteria.

n Penicillin mass-produced in microbes.

1944

n DNA is proven to carry genetic inormationAvery et al.

n Waksman isolates streptomycin, an eective antibioticor tuberculosis.

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8


1946

n Discovery that genetic material rom dierent virusescan be combined to orm a new type o virus, an ex-ample o genetic recombination.

n Recognizing the threat posed by loss o genetic diversity,the U.S. Congress provides unds or systematic andextensive plant collection, preservation and introduction.

1947n McClintock discovers transposable elements, or

jumping genes, in corn.

1949

n Pauling shows that sickle cell anemia is a moleculardisease resulting rom a mutation in the protein mol-ecule hemoglobin.

1951n Articial insemination o livestock using rozen semen

is accomplished.

1953n The scientic journalNature publishes James Watson

and Francis Cricks manuscript describing the doublehelical structure o DNA, which marks the beginningo the modern era o genetics.

1955

n An enzyme involved in the synthesis o a nucleic acidis isolated or the rst time.

1956

n Kornberg discovers the enzyme DNA polymerase I,leading to an understanding o how DNA is repli-

cated.

1958n Sickle cell anemia is shown to occur due to a change

o a single amino acid.

n DNA is made in a test tube or the rst time.

1959

n Systemic ungicides are developed. The steps in pro-tein biosynthesis are delineated.

ALSO IN THE 1950s

n Discovery o intererons.

n First synthetic antibiotic.

1960

n Exploiting base pairing, hybrid DNA-RNA moleculesare created.

n Messenger RNA is discovered.

1961

n

USDA registers rst biopesticide:Bacillus thuringien-sis, or Bt.

1963

n New wheat varieties developed by Norman Borlaugincrease yields by 70 percent.

1964

n The International Rice Research Institute in the Phil-ippines starts the Green Revolution with new strainso rice that double the yield o previous strains i givensucient ertilizer.

1965n Harris and Watkins successully use mouse and human

cells.

1966n The genetic code is cracked, demonstrating that a se-

quence o three nucleotide bases (a codon) determineseach o 20 amino acids. (Two more amino acids havesince been discovered.)

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1967

n The rst automatic protein sequencer is perected.

1969

n An enzyme is synthesized in vitro or the rst time.

1970

n Norman Borlaug receives the Nobel Peace Prize (see1963).

n Discovery o restriction enzymes that cut and splicegenetic material, opening the way or gene cloning.

1971n First complete synthesis o a gene.

1972n The DNA composition o humans is discovered to be

99 percent similar to that o chimpanzees and gorillas.

n Initial work with embryo transer.

1973n Stanley Cohen and Herbert Boyer perect techniques

to cut and paste DNA (using restriction enzymes andligases) and reproduce the new DNA in bacteria.

1974

n The National Institutes o Health orms a Recombi-nant DNA Advisory Committee to oversee recombinantgenetic research.

1975

n Government rst urged to develop guidelines orregulating experiments in recombinant DNA: AsilomarConerence, Caliornia.

n The rst monoclonal antibodies are produced.

1976

n The tools o recombinant DNA are rst applied to ahuman inherited disorder.

n Molecular hybridization is used or the prenatal diag-

nosis o alpha thalassemia.

n Yeast genes are expressed inE. colibacteria.

n The sequence o DNA base pairs or a specic gene isdetermined.

n First guidelines or recombinant DNA experimentsreleased: National Institutes o HealthRecombinantDNA Advisory Committee.

1977

n First expression o human gene in bacteria.

n Procedures developed or rapidly sequencing long sec-tions o DNA using electrophoresis.

1978n High-level structure o virus rst identied.

n Recombinant human insulin rst produced.

n North Carolina scientists show it is possible to in-

troduce specic mutations at specic sites in a DNAmolecule.

1979

n Human growth hormone rst synthesized.

ALSO IN THE 1970s

n First commercial company ounded to develop geneti-cally engineered products.

n Discovery o polymerases.

n

Techniques or rapid sequencing o nucleotides per-ected.

n Gene targeting.

n RNA splicing.

1980n The U.S. Supreme Court, in the landmark case

Diamond v. Chakrabarty, approves the principle opatenting organisms, which allows the Exxon oilcompany to patent an oil-eating microorganism.

n The U.S. patent or gene cloning is awarded to Cohenand Boyer.

n The rst gene-synthesizing machines are developed.

n Researchers successully introduce a human geneone that codes or the protein intereroninto abacterium.

n Nobel Prize in Chemistry awarded or creationo the rst recombinant molecule: Berg, Gilbert,Sanger.

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0


1981n Scientists at Ohio University produce the rst trans-

genic animals by transerring genes rom other ani-mals into mice.

n Chinese scientist becomes the rst to clone a shagolden carp.

1982

n Applied Biosystems, Inc., introduces the rst commer-cial gas phase protein sequencer, dramatically reducingthe amount o protein sample needed or sequencing.

n First recombinant DNA vaccine or livestock developed.

n First biotech drug approved by FDA: human insulinproduced in genetically modied bacteria.

n First genetic transormation o a plant cell: petunia.

1983n The polymerase chain reaction (PCR) technique is

conceived. PCR, which uses heat and enzymes to makeunlimited copies o genes and gene ragments, laterbecomes a major tool in biotech research and productdevelopment worldwide.

n The rst genetic transormation o plant cells by TIplasmids is perormed.

n The rst articial chromosome is synthesized.

n The rst genetic markers or specic inherited dis-eases are ound.

n

First whole plant grown rom biotechnology: petunia.n First proo that modied plants pass their new traits to

ospring: petunia.

1984n The DNA ngerprinting technique is developed.

n The entire genome o the human immunodeciencyvirus is cloned and sequenced.

1985

n Genetic markers ound or kidney disease and cystic

brosis.

n Genetic ngerprinting entered as evidence in a court-room.

n Transgenic plants resistant to insects, viruses andbacteria are eld-tested or the rst time.

n The NIH approves guidelines or perorming gene-therapy experiments in humans.

1986n First recombinant vaccine or humans: hepatitis B.

n First anticancer drug produced through biotech: inter-eron.

n The U.S. government publishes the CoordinatedFramework or Regulation o Biotechnology, estab-lishing more stringent regulations or rDNA organ-isms than or those produced with traditional geneticmodication techniques.

n A University o CaliorniaBerkeley chemist describeshow to combine antibodies and enzymes (abzymes) tocreate pharmaceuticals.

n The rst eld tests o transgenic plants (tobacco) areconducted.

n The Environmental Protection Agency approves therelease o the rst transgenic cropgene-alteredtobacco plants.

n The Organization o Economic Cooperation and Devel-opment (OECD) Group o National Experts on Saety inBiotechnology states: Genetic changes rom rDNA tech-niques will oten have inherently greater predictability

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compared to traditional techniques and risks associatedwith rDNA organisms may be assessed in generally thesame way as those associated with non-rDNA organisms.

1987

n First approval or eld test o modied ood plants:virus-resistant tomatoes.

n Frostban, a genetically altered bacterium that inhibitsrost ormation on crop plants, is eld-tested onstrawberry and potato plants in Caliornia, the rstauthorized outdoor tests o a recombinant bacterium.

1988

n Harvard molecular geneticists are awarded the rst U.S.

patent or a genetically altered animala transgenicmouse.

n A patent or a process to make bleach-resistant prote-ase enzymes to use in detergents is awarded.

n Congress unds the Human Genome Project, a massiveeort to map and sequence the human genetic code as

well as the genomes o other species.

1989n First approval or eld test o modied cotton: insect-

protected (Bt) cotton.

n Plant Genome Project begins.

ALSO IN THE 1980s

n Studies o DNA used to determine evolutionary history.

n Recombinant DNA animal vaccine approved or use inEurope.

n Use o microbes in oil spill cleanup: bioremediationtechnology.

n Ribozymes and retinoblastomas identied.

1990n Chy-Max, an articially produced orm o the chymo-

sin enzyme or cheese-making, is introduced. It is therst product o recombinant DNA technology in theU.S. ood supply.

n The Human Genome Projectan international eortto map all the genes in the human bodyis launched.

n The rst experimental gene therapy treatment is per-ormed successully on a 4-year-old girl suering roman immune disorder.

n The rst transgenic dairy cowused to produce hu-man milk proteins or inant ormulais created.

n First insect-protected corn: Bt corn.n First ood product o biotechnology approved in U.K.:

modied yeast.

n First eld test o a genetically modied vertebrate: trout.

1992

n American and British scientists unveil a technique ortesting embryos in vitro or genetic abnormalities suchas cystic brosis and hemophilia.

n The FDA declares that transgenic oods are not inher-

ently dangerous and do not require special regulation.

1993n Merging two smaller trade associations creates the

Biotechnology Industry Organization (BIO).

n FDA approves bovine somatotropin (BST) or increasedmilk production in dairy cows.

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2


1994

n First FDA approval or a whole ood producedthrough biotechnology: FLAVRSAVR tomato.

n The rst breast cancer gene is discovered.

n Approval o recombinant version o human DNase,which breaks down protein accumulation in the lungso CF patients.

n BST commercialized as POSILAC bovine somatotropin.

1995

n The rst baboon-to-human bone marrow transplant isperormed on an AIDS patient.

n The rst ull gene sequence o a living organism otherthan a virus is completed, or the bacteriumHemophi-lus infuenzae.

n Gene therapy, immune system modulation and recom-binantly produced antibodies enter the clinic in the

war against cancer.

1996

n The discovery o a gene associated with Parkinsonsdisease provides an important new avenue o researchinto the cause and potential treatment o the debilitat-ing neurological ailment.

1997

n First animal cloned rom an adult cell: a sheep namedDolly in Scotland.

n First weed- and insect-resistant biotech crops com-mercialized: Roundup Ready soybeans and Bollgardinsect-protected cotton.

n Biotech crops grown commercially on nearly 5 millionacres worldwide: Argentina, Australia, Canada, China,Mexico and the United States.

n A group o Oregon researchers claims to have clonedtwo Rhesus monkeys.

1998n University o Hawaii scientists clone three generations

o mice rom nuclei o adult ovarian cumulus cells.

n Human embryonic stem cell lines are established.

n Scientists at Japans Kinki University clone eight iden-tical calves using cells taken rom a single adult cow.

n The rst complete animal genome, or the C. elegansworm, is sequenced.

n

A rough drat o the human genome map is produced,showing the locations o thousands o genes.

n Five Southeast Asian countries orm a consortium todevelop disease-resistant papayas.

ALSO IN THE 1990sn First conviction using genetic ngerprinting in the U.K.

n Discovery that hereditary colon cancer is caused bydeective DNA repair gene.

n Recombinant rabies vaccine tested in raccoons.

n Biotechnology-based biopesticide approved or sale inthe United States.

n Patents issued or mice with specic transplantedgenes.

n First European patent on a transgenic animal issuedor transgenic mouse sensitive to carcinogens.

2000

n First complete map o a plant genome developed:Ara-bidopsis thaliana.

n Biotech crops grown on 108.9 million acres in 13countries.

n Golden rice announcement allows the technologyto be available to developing countries in hopes oimproving the health o undernourished people andpreventing some orms o blindness.

n First biotech crop eld-tested in Kenya: virus-resistantsweet potato.

n Rough drat o the human genome sequence is an-nounced.

2001

n First complete map o the genome o a ood plantcompleted: rice.

n Researchers with Chinas National Hybrid Rice Re-search Center report developing a super rice thatcould produce double the yield o normal rice.

n Complete DNA sequencing o the agriculturallyimportant bacteriaSinorhizobium meliloti, a nitro-

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gen-xing species, andAgrobacterium tumeaciens, aplant pest.

n

A single gene romArabidopsis inserted into tomatoplants to create the rst crop able to grow in salty

water and soil.

2002n The rst drat o a unctional map o the yeast pro-

teome, an entire network o protein complexes andtheir interactions, is completed. A map o the yeastgenome was published in 1996.

n International consortia sequence the genomes o theparasite that causes malaria and the species o mos-quito that transmits the parasite.

n The drat version o the complete map o the humangenome is published, and the rst part o the HumanGenome Project comes to an end ahead o scheduleand under budget.

n Scientists make great progress in elucidating the ac-tors that control the dierentiation o stem cells, iden-tiying over 200 genes that are involved in the process.

n Biotech crops grown on 145 million acres in 16 coun-tries, a 12 percent increase in acreage. More than one-quarter (27 percent) o the global acreage was grown

in nine developing countries.

n Researchers announce successul results or a vaccineagainst cervical cancer, the rst demonstration o a

preventative vaccine or a type o cancer.

n Scientists complete the drat sequence o the most im-portant pathogen o rice, a ungus that destroys enoughrice to eed 60 million people annually. By combining anunderstanding o the genomes o the ungus and rice,scientists can elucidate the molecular basis o the inter-actions between the plant and pathogen.

n

Scientists are orced to rethink their view o RNAwhen they discover how important small pieces oRNA are in controlling many cell unctions.

n Japanese puersh genome is sequenced. The pu-ersh sequence is the smallest known genome o any

vertebrate.

n Scientists at Stony Brook University in New Yorkassemble a synthetic virus, polio, using genomesequence inormation. The project raises ethical andsecurity questions.

2003n Researchers nd a vulnerability gene or depression

and make strides in detecting genetic links to schizo-phrenia and bipolar disorder.

n GloFish, the rst biotech pet, hits the North Ameri-can market. Specially bred to detect water pollutants,the sh glows red under black light thanks to the addi-tion o a natural fuorescence gene.

n Worldwide biotech crop acreage rises 15 percent to hit167.2 million acres in 18 countries. Brazil and the Philip-pines grow biotech crops or the rst time in 2003. Also,Indonesia allows consumption o imported biotech oods,

and China and Uganda accept biotech crop imports.

n The U.K. approves its rst commercial biotech crop ineight years. The crop is a biotech herbicide-resistantcorn used or cattle eed.

n The U.S. Environmental Protection Agency approvesthe rst transgenic rootworm-resistant corn, whichmay save armers $1 billion annually in crop lossesand pesticide use.

n An endangered species (the banteng) is cloned or therst time. 2003 also brought several other cloning rsts,including mules, horses and deer.

n Dolly, the cloned sheep that made headlines in 1997, iseuthanized ater developing progressive lung disease.Dolly was the rst successul clone o an adult mammal.

n Japanese researchers develop a biotech coee beanthat is naturally decaeinated.

n Chinas State Food and Drug Administration grantsthe worlds rst regulatory approval o a gene therapyproduct, Gendicine, developed by Shenzhen SiBionoGenTech. The product delivers the p53 gene as a thera-py or squamous cell head and neck cancer.

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2004

n The FDA approves the rst anti-angiogenic drug orcancer, AVASTIN (bevacizumab).

n The FDA clears a DNA microarray test system, theAmpliChip Cytochrome P450 Genotyping Test, to aidin selecting medications or a wide variety o common

conditions.

n An RNA-intererence product or age-related wetmacular degeneration becomes the rst RNAi productto enter a clinical trial.

n The United Nations Food and Agriculture Organiza-tion endorses biotech crops and states that biotechnol-ogy is a complementary tool to traditional arming

methods that can help poor armers and consumers indeveloping nations.

n The National Academy o Sciences Institute o Medicinends biotech crops do not pose any more health risksthan do crops created by other techniques, and that oodsaety evaluations should be based on the resulting oodproduct, not the technique used to create it.

n FDA nds biotech wheat sae ater a ood saety review.

n Monsanto introduces low-linolenic soybeans (pro-duced through conventional breeding methods) that

will reduce or eliminate trans atty acids in processed

soybean oil.

n Chicken genome sequenced by the Chicken GenomeSequencing Consortium.

n First cloned pet, a kitten, is delivered to its owner.

n Laboratory rat genome is sequenced.

n Researchers complete the sequence o the chimpan-zeehumanitys closest primate relative.

n The Canadian biotech company Iogen achieves the rstcommercial production and delivery o bioethanol, pro-

ducing the uel with biotech enzymes and wheat straw.

n Caliornia voters pass Proposition 71, which supportsembryonic stem cell research with $3 billion in und-ing over 10 years.

2005

n Researchers at the University o Georgia successullyproduce a cow cloned rom the cells o a carcass.

n FDA or the rst time approves a drug or a specicrace. The drug, NitroMeds BiDil, treats congestive

heart ailure in sel-identied black patients. Thecompany hopes a genetic test can be developed toidentiy patients likely to benet, regardless o race.

n The Energy Policy Act is passed and signed into law,authorizing numerous incentives or bioethanol develop-ment.

n The National Institutes o Health in December launch-es a pilot project to determine the easibility o TheCancer Genome Atlas. The ultimate goal would be acomplete map o the genomic changes involved in alltypes o human cancer.

n Using new genome sequence inormation, scientists atthe Centers or Disease Control & Prevention partially

synthesize the fu virus that killed at least 20 millionpeople worldwide in 19181919.

n Scientists at Harvard University report success in con-verting skin cells into embryonic stem cells throughusion with existing embryonic stem cells.

n USDA, Monsanto and Genaissance Pharmaceuticalsannounce a joint soybean genome project.

n The British government approves the Equine Fertil-ity Units research in using nuclear transer in horsecloning.

n On May 7, the one billionth acre o biotech seed isplanted.

n The World Health Organization (WHO) issues the reportModern Food Biotechnology, Human Health and De-velopment, which states biotech oods can contribute toenhancing human health and development. Accordingto the report, biotech oods can increase crop yield, oodquality, and the diversity o oods which can be grown ina given area. They lead to better health and nutrition andthereby help raise health and living standards.

n The British research rm PG Economics Ltd. nds

that the global use o biotech crops has added $27 bil-lion to arm income, and greatly reduced agriculturesnegative impacts on the environment.

n The National Science Foundation, USDA, and theDepartment o Energy award $32 million to a teamo university and private laboratory researchers tosequence the corn genome.

n A consortium o scientists led by the National HumanGenome Research Institute publishes the dog genome,

which belongs to a 12-year-old boxer.

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n Global biotech crop acreage reaches 222 million acres.

2006

n A team o researchers headed by the University o Il-linois receives a $10 million ederal grant to completethe sequence o the swine genome. The project isexpected to be completed within two years.

n In his State o the Union address, President Bushexpresses support or bioethanol made rom agricul-tural wastes. Doesnt it make sense, he asked, todetermine whether or not we can use these raw ma-terials to make something out o nothing so that wecontinue the advance o ethanol and so the marketor ethanol expands throughout the United States?

n The National Institutes o Health begins a 10-year,10,000-patient study using a genetic test that pre-dicts breast-cancer recurrence and guides treatment.Patients whose cancer is deemed unlikely to recur willbe spared chemotherapy. The genetic test, OncotypeDX was developed by the biotech company GenomicHealth and is already commercially available.

n In January, the American Dietetic Association (ADA)publishes a rearmed statement o support on agri-cultural and ood biotechnology. The ADA states thatagricultural biotechnology techniques can enhance

the quality, saety, nutritional value, and variety o oodavailable or human consumption while increasing theeciency o ood production, ood processing, ood dis-tribution, and environmental and waste management.

n Dow AgroSciences announces it has received the rstregulatory approval or a plant-made vaccine romUSDAs Center or Veterinary Biologics. The vaccineprotects poultry rom Newcastle disease, and is therst plant-made vaccine to be approved.

n Renessen LLC, a joint venture o Monsanto and Cargill,receives approval rom USDA to begin selling the rst

crop improved through biotechnology with added ben-ets or use in animal eed. The product, Mavera HighValue Corn with Lysine, has been improved to grow withincreased levels o lysine, an amino acid that is essentialor animal diets, especially those o swine and poultry.

n USDA awards $5 million to a consortium o public wheatbreeders and 18 universities or wheat genome research.

n Researchers develop biotech pigs that produce highlevels o omega-3 atty acids. The biotech pigs were de-

veloped by inserting the at-1 gene that comes rom

the roundworm Caenorhabditis elegans. The biotechpigs were cloned, and six o the 10 clones producedincreased levels o omega-3 atty acids, which are

believed to ward o heart disease.

n The World Trade Organization issues a condentialnal ruling on the U.S./Canada/Argentine challengeagainst the European Union (EU) on approval onew biotech crops. According to news reports, theruling concludes that the EU breached its tradecommitments with respect to 21 agricultural bio-technology productsincluding types o oilseed,rape, maize and cotton.

n The French agriculture ministry authorizes 17 neweld tests or biotech corn and tobacco crops.

Sources:Access ExcellenceBiotech 90: Into the Next Decade,

G. Steven Burrill with the Ernst & Young HighTechnology Group

Biotechnology Industry OrganizationGenentech, Inc.Genetic Engineering NewsInternational Food Inormation Council

ISB News ReportInternational Service or the

Acquisition o Agri-BiotechApplications

Texas Society or Biomedical ResearchScienceScience NewsThe Scientist

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Biotechnology Policy Milestones

1902n The Biologics Control Act passes to ensure purity and

saety o serums, vaccines and similar products.

1906

n The Food and Drugs Act is signed into law, prohibitinginterstate commerce in misbranded and adulteratedood, drinks and drugs. (Note: For a detailed FDAtimeline, visit http://www.da.gov/opacom/background-ers/miles.html.)

1930n The National Institute o Health is created (later to be

pluralized as National Institutes o Health).

1971n President Nixon calls or a War on Cancer and signs

the National Cancer Act into law, stimulating newresearch.

1974n Leading biologists call or a voluntary moratorium on

recombinant DNA experiments while saety standards

are set.

1975n Some 150 scientists, attorneys, government ocials

and journalists meet at the Asilomar Conerence Cen-ter near Monterey, Cali., to discuss recombinant DNAresearch and develop strict saety protocols.

1976

n The NIH adopts guidelines or ederally unded recom-binant DNA research, with oversight provided by theRecombinant DNA Advisory Committee.

1980

n The Supreme Court decides inDiamond vs. Chakrab-arty that anything made by the hand o man, includ-ing biotechnology-modied organisms, is patentable.The decision helps open the foodgates to a wave oinvestment that includes the rst biotech IPOs.

n The Patent and Trademark Act Amendments o1980commonly known as the Bayh-Dole Actlay

the ground rules or technology transer rom aca-demia to industry. The act creates a uniorm patentpolicy among ederal agencies that und research and

species that ederal grant recipientssuch as uni-versities and small businessesown ederally undedinventions.

1983n The Orphan Drug Act is signed into law, creating new

incentives to conduct R&D on therapies or rare dis-eases. More than 250 orphan drugs have reached theU.S. market in the years since.

1988

n The U.S. Patent and Trademark Oce grants Har-vard University a patent or a mouse used or cancerresearch (the OncoMouse).

n The United States launches the Human GenomeProject when Congress appropriates unds or theDepartment o Energy and the National Institutes oHealth to support research to determine the structureo complex genomes.

1992n The FDA clears the way or agricultural biotechnol-

ogy products with a saety assessment and guidance to

industry.

n The Prescription Drug User Fee Act is signed into law,instituting ees or drug approval applicants that pro-

vide the FDA with resources to review products aster.The successul program is reauthorized in 1997 and2002 and is up or reauthorization again in 2007.

1993n The Biotechnology Industry Organization (BIO) is

created out o the merger o two predecessor organiza-tions, the Industrial Biotechnology Association and the

Association o Biotechnology Companies. (A history oBIO is posted on bio.org in the About BIO section.)

1997

n The Food and Drug Administration Modernization Act(FDAMA) is signed into law, codiying administrativechanges begun in 1995 and introducing new reorms.Provisions include criteria or ast-track drug develop-ment, easier patient access to experimental drugs andmedical devices, and an online database o clinical trials.

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1998

n Congress undertakes a doubling o the National Insti-tutes o Health budget in ve years, raising it to $27

billion by 2003.

2000n The Biomass Research and Development Act is signed

into law to promote conversion o biomass into bio-based industrial products.

2001n President Bush announces that ederal unding will be

made available to support research using embryonicstem cell lines created as o Aug. 9, 2001.

2002

n The Farm Security and Rural Investment Act includesbiotech measures such as signicantly increased und-ing or research and risk assessment and new programsor promoting biotechnology in developing countries.

2003

n The Medicare Modernization Act becomes law, provid-ing prescription drug coverage or senior citizens andthe disabled beginning Jan. 1, 2006.

2004n The FDA publishes a white paper outlining the

Critical Path Initiative, which seeks to expedite drugdevelopment by promoting the use o technologiessuch as computer-based predictive models, biomark-ers, imaging technologies and improved clinicaltrial design.

n The Project BioShield Act is signed into law, providing$5.6 billion over 10 years or the ederal governmentto procure diagnostics, therapies and vaccines to pro-tect Americans rom chemical, nuclear and biological

warare agents.

2005n The Energy Policy Act o 2005 passes, authorizing

$3.6 billion in unding or bioenergy and biobasedproducts.

n Pandemic legislation signed into law provides $3.8 bil-lion or preparedness, including $3 billion or medicalcountermeasures. The legislation also includes liabilityprotection or manuacturers o these products.

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H

ere are a ew o the new biotechnologies that usecells and biological molecules and examples otheir applications in medicine, agriculture, ood

processing, industrial manuacturing and environmentalmanagement.

Bioprocessing Technology

The oldest o the biotechnologies, bioprocessingtechnology, uses living cells or the molecular com-ponents o their manuacturing machinery to producedesired products. The living cells most commonly usedare one-celled microorganisms, such as yeast and bacte-ria; the biomolecular components we use most oten are

enzymes, which are proteins that catalyze biochemicalreactions.

A orm o bioprocessing,microbial ermentation, hasbeen used or thousands o yearsunwittinglytobrew beer, make wine, leaven bread and pickle oods.In the mid-1800s, when we discovered microorganismsand realized their biochemical machinery was respon-sible or these useul products, we greatly extended ouruse o microbial ermentation. We now rely on the re-markably diverse manuacturing capability o naturallyoccurring microorganisms to provide us with productssuch as antibiotics, birth control pills, amino acids,

vitamins, industrial solvents, pigments, pesticides andood-processing aids.

Today, we are using recombinant DNA technology,coupled with microbial ermentation, to manuacture a

wide range o biobased products including human insulin,the hepatitis B vaccine, the cal enzyme used in cheese-making, biodegradable plastics, and laundry detergentenzymes. Bioprocessing technology also encompassestissue engineering and manuacturing as well as biophar-maceutical ormulation and delivery.

Monoclonal Antibodies

Monoclonal antibody technology uses immune-systemcells that make proteins called antibodies. We haveall experienced the extraordinary specicity o antibod-ies: Those that attack a fu virus one winter do nothing toprotect us rom a slightly dierent fu virus the next year.(Specicity reers to the act that biological molecules aredesigned so that they bind to only one molecule.)

The specicity o antibodies also makes them poweruldiagnostic tools. They can locate substances that occurin minuscule amounts and measure them with great ac-

curacy. For example, we use monoclonal antibodies to

n locate environmental pollutants.

n detect harmul miroorganisms in ood.

n distinguish cancer cells rom normal cells.

n diagnose inectious diseases in humans, animals andplants more quickly and more accurately than everbeore.

In addition to their value as detection devices, monoclo-nal antibodies (MAbs) can provide us with highly specic

therapeutic compounds. Monoclonal antibodies joined to atoxin can selectively deliver chemotherapy to a cancer cell

while avoiding healthy cells. We are developing monoclonalantibodies to treat organ-transplant rejection and autoim-mune diseases by targeting them specically to the type oimmune system cell responsible or these attacks, leavingintact the other branches o the immune system.

MAbs FOR IMMUNE-RELATED CONDITIONS

n Muromomab-CD3 (OKT3) is used to prevent acuterejection o organ transplants. A modied version oOKT3 shows promise in inhibiting the autoimmune

destruction o beta cells in Type 1 diabetes mellitus.

n Infiximab (Remicade) binds to tumor necrosis ac-tor-alpha and has shown promise against some infam-matory diseases such as rheumatoid arthritis.

n Omalizumab (Xolair) binds to IgE and prevents itrom binging to mast cells. The drug is used againstallergic asthma.

n Daclizumab (Zenapax) binds to part o the IL-2 re-ceptor and is used to prevent acute rejection o trans-planted kidneys. The drug also shows promise againstT-cell lymphoma.

MAbs USED TO KILL OR INHIBIT CANCER CELLS

n Rituximab (Rituxan) binds to the CD20 molecule thatis ound on most B-cells and is used to treat B-cell lym-phomas

n Ibritumomab tiuxetan (Zevalin) is used against theCD20 molecule on B-cells (and lymphomas) conjugat-ed to either o two radioactive isotopes in conjunction

with Rituxan.

The Technologies and Their Applications

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n Tositumomab (Bexxar) is a conjugate o a monoclo-nal antibody against CD20 and the radioactive isotopeiodine-131. It has been approved to treat lymphoma.

n Trastuzumab (Herceptin) binds to HER2, a recep-tor or epidermal growth actor ound on some breastcancers and lymphomas.

n Cetuximab (Erbitux) blocks HER1, another epider-mal growth actor receptor, and has been approved totreat colorectal cancer.

n Gemtuzumab ozogamicin (Mylotarg) is a conjugateo a monoclonal antibody that binds to CD33, a cell-surace molecule expressed by the cancerous cells inacute myelogenous leukemia, and calicheamicin, a

complex oligosaccharide that makes double-strandedbreaks in DNA. The drug is the rst immunotoxin thatshows promise in the ght against cancer.

n Alemtuzumab (Campath) binds to CD52, a moleculeound on white blood cells, and has produced completeremission o chronic lymphocytic leukemia (or 18months and counting).

ANGIOGENESIS INHIBITOR

n Bevacizumab (Avastin) blocks the vascular endo-thelial growth actor (VEGF) receptor and has beenapproved or the treatment o colorectal cancer.

OTHER

n Abciximab (ReoPro) inhibits the clumping o plate-lets by binding the receptors on their surace that nor-mally are linked by brinogen. This therapy is helpulin preventing the re-clogging o the coronary arteriesin patients who have undergone angioplasty.

Monoclonal antibodies can be created in mice, but mouseantibodies are seen by the human immune system andoten the human patient mounts an immune response,

which not only eliminates the therapeutic MAb adminis-

tered, but also causes damage to the kidneys. To reducethe problem o human anti-mouse antibodies (HAMA),scientists use chimeric, or humanized, antibodies. Toorm a chimeric antibody, one must combine the antigen-binding parts (variable regions) o the mouse antibody

with the eector parts (constant regions) o a humanantibody. Infiximab, rituximab and abciximab are exam-ples. To create human antibodies, one combines only theamino acids responsible or making the antigen bindingsite (the hypervariable regions) o a mouse antibody and

the rest o a human antibody molecule, thus replacing itsown hypervariable regions. Zenapax, Vitaxin, Mylotarg,Herceptin, and Xolair are examples.

Cell Culture

C ell culture technology is the growing o cells outsideo living organisms.PLANT CELL CULTURE

An essential step in creating transgenic crops, plant cellculture also provides us with an environmentally soundand economically easible option or obtaining naturallyoccurring products with therapeutic value, such as the

chemotherapeutic agent paclitaxel, a compound oundin yew trees and marketed under the name Taxol. Plantcell culture is also an important source o compoundsused as favors, colors and aromas by the ood-processingindustry.

INSECT CELL CULTURE

Insect cell culture can broaden our use o biological con-trol agents that kill insect pests without harming benecialinsects or having pesticides accumulate in the environ-ment. Even though we have recognized the environmen-

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tal advantages o biological control or many decades,manuacturing biological control products in marketableamounts has been impossible. Insect cell culture removes

these manuacturing constraints. In addition, like plant cellculture, insect cell culture is being investigated as a pro-duction method o therapeutic proteins. Insect cell cultureis also being investigated or the production o VLP (virus-like particle) vaccines against inectious diseases such asSARS and infuenza, which could lower costs and eliminatethe saety concerns associated with the traditional egg-based process. A patient specic cancer vaccine that utilizesinsect cell culture has reached Phase III clinical trials.

MAMMALIAN CELL CULTURE

Livestock breeding has used mammalian cell culture as

an essential tool or decades. Eggs and sperm, taken romgenetically superior bulls and cows, are united in the lab,and the resulting embryos are grown in culture beore be-ing implanted in surrogate cows. A similar orm o mam-malian cell culture has also been an essential componento the human in vitro ertilization process.

Our use o mammalian cell culture now extends wellbeyond the brie maintenance o cells in culture orreproductive purposes. Mammalian cell culture cansupplementand may one day replaceanimal testingto assess the saety and ecacy o medicines. Like plant

cell culture and insect cell culture, we are relying on themanuacturing capacity o mammalian cells to synthesizetherapeutic compounds, in particular, certain mammalianproteins too complex to be manuactured by geneticallymodied microorganisms. For example, monoclonal anti-bodies are produced through mammalian cell culture.

Scientists are also investigating the use o mammaliancell culture as a production technology or vaccines. In2005, the Department o Health and Human Servicesawarded a $97 million contract to Sano Pasteur todevelop mammalian cell culturing techniques to speedthe production process or new infuenza vaccines and

thereby enhance pandemic preparedness.

Therapies based on culturedadult stem cells, which areound in certain tissues like the bone marrow and brain,are on the horizon as well. Researchers have ound thatadult stem cells can be used by the body to replenishtissues. Adult hematopoietic stem cells already are beingtransplanted into bone marrow to stimulate the gen-eration o the various types o blood cells necessary torejuvenate an immune system. These stem cells can beharvested in large quantities rom umbilical cord blood,but they are dicult to isolate and puriy.

Researchers also are working on ways to harvest stemcells rom placentas and rom at. Some are looking atcellular reprogramming as a way to get specialized body

cells, like skin cells, to revert to a primordial state so thatthey can be coaxed into various types o tissues.

Embryonic stem cells are also under study as potentialtherapies. As the name suggests, embryonic stem cells arederived rom embryosspecically those that develop romeggs that have been ertilized in vitro (in an in vitro ertiliza-tion clinic) and then donated by consent or research pur-poses. The embryos are typically our or ve days old and areeach a hollow microscopic ball o cells called the blastocyst.

The potential value of stem cell therapy and tissue

engineering can best be realized if the therapeutic

stem cells and the tissues derived from them aregenetically identical to the patient receiving them.

Human embryonic stem cells are isolated by transerringthe inner cell mass into a nutrient rich culture medium.There the human stem cells prolierate. Over the courseo several days, the cells o the inner cell mass divide andspread all over the dish. Researchers then must removethe growing cells and divide them into resh culturedishes. This process o replating the cells, called subcul-turing, is repeated many times over many months. Eachcycle o subculturing cells is called apassage. Embryonic

stem cells that have prolierated in cell culture or six ormore months without dierentiating (i.e., remain plu-ripotent) and appear genetically normal are reerred to asanembryonic stem cell line.

The inner surace o the culture dish may be coated withmouse embryonic skin cells that have been engineerednot to divide. This is called the eeder layer. It provides asticky surace to which the human embryonic cells attach.Recently scientists have been guring out ways to growembryonic stem cells without using mouse eeder cellsasignicant advance because o the risk o viruses andother macromolecules in the mouse cells being transmit-

ted to the human cells.

The potential value o stem cell therapy and tissue engi-neering can best be realized i the therapeutic stem cellsand the tissues derived rom them are genetically identicalto the patient receiving them. Thereore, unless the patientis the source o the stem cells, the stem cells need to becustomized by replacing the stem cells genetic material

with the patients beore cueing the stem cells to dierenti-ate into a specic cell type. To date, this genetic materialreplacement and reprogramming can be done eectivelyonly with embryonic stem cells.

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Recombinant DNA Technology

Recombinant DNA technology is viewed by many as thecornerstone o biotechnology. The termrecombinant

DNA literally means the joining or recombining o twopieces o DNA rom two dierent species.

Humans began to preerentially combine the geneticmaterial o domesticated plants and animals thousands o

years ago by selecting which individuals would reproduce.By breeding individuals with valuable genetic traits whileexcluding others rom reproduction, we changed the ge-netic makeup o the plants and animals we domesticated.Now, in addition to using selective breeding to combine

valuable genetic material rom dierent organisms, we

combine genes at the molecular level using the moreprecise techniques o recombinant DNA technology.

Genetic modication through selective breeding and re-combinant DNA techniques undamentally resemble eachother, but there are important dierences:

n Genetic modication using recombinant DNA tech-niques allows us to move single genes whose unctions

we know rom one organism to any other.

n In selective breeding, large sets o genes o unknownunction are transerred between related organisms.

By making our manipulations more precise and ouroutcomes more certain, we decrease the risk o producingorganisms with unexpected traits and avoid the time-con-suming, trial-and-error approach o selective breeding. Byincreasing the breadth o species rom which we can obtainuseul genes, we can access all o natures genetic diversity.

Techniques or making selective breeding more predictableand precise have been evolving over the years. In the early1900s, Hugo DeVries, Karl Correns and Eric Tshermarkrediscovered Mendels laws o heredity. In 1953, James

Watson and Francis Crick deduced DNAs structure rom

experimental clues and model building. In 1972, PaulBerg and colleagues created the rst recombinant DNAmolecules, using restriction enzymes. Ten years later, therst recombinant DNA-based drug (recombinant humaninsulin) was introduced to the market. By 2000 the humangenome had been sequenced and today we use recombinantDNA techniques, in conjunction with molecular cloning to

n produce new medicines and saer vaccines.

n treat some genetic diseases.

n enhance biocontrol agents in agriculture.

n increase agricultural yields and decrease productioncosts.

n

decrease allergy-producing characteristics o someoods.

n improve oods nutritional value.

n develop biodegradable plastics.

n decrease water and air pollution.

n slow ood spoilage.

n control viral diseases.

n inhibit infammation.

Cloning

Cloning technology allows us to generate a population ogenetically identical molecules, cells, plants or animals.Because cloning technology can be used to produce mol-ecules, cells, plants and some animals, its applications areextraordinarily broad. Any legislative or regulatory actiondirected at cloning must take great care in dening theterm precisely so that the intended activities and productsare covered while others are not inadvertently captured.

MOLECULAR OR GENE CLONINGMolecular or gene cloning, the process o creating geneti-cally identical DNA molecules, provides the oundationo the molecular biology revolution and is a undamentaland essential tool o biotechnology research, developmentand commercialization. Virtually all applications in bio-technology, rom drug discovery and development to theproduction o transgenic crops, depend on gene cloning.

Virtually all applications in biotechnology, from drug

discovery and development to the production oftransgenic crops, depend on gene cloning.

The research ndings made possible through molecularcloning include identiying, localizing and characteriz-ing genes; creating genetic maps and sequencing entiregenomes; associating genes with traits and determiningthe molecular basis o the trait. For a ull discussion,see page 27.

ANIMAL CLONING

Animal cloning has helped us to rapidly incorporate im-provements into livestock herds or more than two decadesand has been an important tool or scientic researchers

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since the 1950s. Although the 1997 debut o Dolly, thecloned sheep, brought animal cloning into the public con-sciousness, the production o an animal clone was not a new

development. Dolly was considered a scientic breakthroughnot because she was a clone, but because the source o thegenetic material that was used to produce Dolly was an adultcell, not an embryonic one.

Recombinant DNA technologies, in conjunction withanimal cloning, are providing us with excellent animalmodels or studying genetic diseases, aging and cancerand, in the uture, will help us discover drugs and evalu-ate other orms o therapy, such as gene and cell therapy.

Animal cloning also provides zoo researchers with a toolor helping to save endangered species.

There are two dierent ways to make an exact genetic copyo an organism such as a sheep or a laboratory mouse.

Articial embryo twinning (AET) is the old-ashionedway to clone. AET mimics the natural process o creat-ing identical twins, only in a Petri dish rather than themothers womb. Researchers manually separate a veryearly embryo into individual cells and then allow each cellto divide and develop on its own. The resulting embryosare placed into a surrogate mother, where they are carriedto term and delivered. Since all the embryos come romthe same zygote, they are genetically identical.

Somatic cell nuclear transer (SCNT) involves the isolationo a somatic (body) cell, which is any cell other then thoseused or reproduction (sperm and egg, known as the germcells). In mammals, every somatic cell has two completesets o chromosomes, whereas the germ cells have onlyone complete set. To make Dolly, scientists transerred thenucleus o a somatic cell taken rom an adult emale sheepand transerred it to an egg cell rom which the nucleus hadbeen removed. Ater some chemical manipulation, the eggcell, with the new nucleus, behaved like a reshly ertilizedzygote. It developed into an embryo, which was implantedinto a surrogate mother and carried to term.

Protein Engineering

Protein engineering technology is used, oten inconjunction with recombinant DNA techniques, toimprove existing proteins, such as enzymes, antibodiesand cell receptors, and to create proteins not ound innature. These proteins may be used in drug develop-ment, ood processing and industrial manuacturing.

The most pervasive uses o protein engineering to dateare applications that alter the catalytic properties oenzymes to develop ecologically sustainable industrial

processes. Enzymes are environmentally superior tomost other catalysts used in industrial manuacturingbecause, as biocatalysts, they dissolve in water and workbest at neutral pH and comparatively low temperatures.In addition, because biocatalysts are more specic thanchemical catalysts, they also produce ewer unwantedbyproducts. The chemical, textile, pharmaceutical, pulpand paper, ood and eed, and energy industries are allbeneting rom cleaner, more energy-ecient produc-tion made possible by incorporating biocatalysts intotheir production processes.

The characteristics that make biocatalysts environmentallyadvantageous may, however, limit their useulness in certainindustrial processes. For example, most enzymes all apartat high temperatures. Scientists are circumventing theselimitations by using protein engineering to increase enzymestability under harsh manuacturing conditions.

In addition to industrial applications, medical research-ers have used protein engineering to design novelproteins that can bind to and deactivate viruses and tu-mor-causing genes; create especially eective vaccines;and study the membrane receptor proteins that are sooten the targets o pharmaceutical compounds. Food

scientists are using protein engineering to improve theunctionality o plant storage proteins and develop newproteins as gelling agents.

In addition, new proteins are being developed to respondto chemical and biological attacks. For example, hydrolas-es detoxiy a variety o nerve agents as well as commonlyused pesticides. Enzymes are sae to produce, store anduse, making them an eective and sustainable approachto toxic materials decontamination.

BiosensorsB iosensor technology couples our knowledge o biol-ogy with advances in microelectronics. A biosensoris composed o a biological component, such as a cell,enzyme or antibody, linked to a tiny transducera devicepowered by one system that then supplies power (usu-ally in another orm) to a second system. Biosensors aredetecting devices that rely on the specicity o cells andmolecules to identiy and measure substances at extreme-ly low concentrations.

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When the substance o interest binds with the biologi-cal component, the transducer produces an electrical oroptical signal proportional to the concentration o the

substance. Biosensors can, or example,

n measure the nutritional value, reshness and saety oood.

n provide emergency room physicians with bedside mea-sures o vital blood components.

n locate and measure environmental pollutants.

n detect and quantiy explosives, toxins and biowarareagents.

Nanobiotechnology

Nanotechnology, which came into its own in 2000with the birth o the National Nanotechnology Ini-tiative, is the next stop in the miniaturization path thatgave us microelectronics, microchips and microcir-cuits. The wordnanotechnology derives romnanome-ter, which is one-thousandth o a micrometer (micron),or the approximate size o a single molecule. Nanotech-nologythe study, manipulation and manuacture oultra-small structures and machines made o as ew asone moleculewas made possible by the development

o microscopic tools or imaging and manipulatingsingle molecules and measuring the electromagneticorces between them.

Nanobiotechnology joins the breakthroughs in nanotech-nology to those in molecular biology. Molecular biologistshelp nanotechnologists understand and access the nano-structures and nanomachines designed by 4 billion yearso evolutionary engineeringcell machinery and biologi-cal molecules. Exploiting the extraordinary properties obiological molecules and cell processes, nanotechnologistscan accomplish many goals that are dicult or impossibleto achieve by other means.

For example, rather than build silicon scaolding or nano-structures, DNAs ladder structure provides nanotechnolo-gists with a natural ramework or assembling nanostruc-tures and its highly specic bonding properties bring atomstogether in a predictable pattern to create a nanostructure.

Nanotechnologists also rely on the sel-assembling prop-erties o biological molecules to create nanostructures,such as lipids that spontaneously orm liquid crystals.

Most appropriately, DNA, the information storagemolecule, may serve as the basis of the nextgeneration of computers.

DNA has been used not only to build nanostructures butalso as an essential component o nanomachines. Mostappropriately, DNA, the inormation storage molecule,may serve as the basis o the next generation o com-puters. As microprocessors and microcircuits shrinkto nanoprocessors and nanocircuits, DNA moleculesmounted onto silicon chips may replace microchips withelectron fow-channels etched in silicon. Such biochipsare DNA-based processors that use DNAs extraordinaryinormation storage capacity. Conceptually, they are verydierent rom the DNA chips discussed below. Biochips

exploit the properties o DNA to solve computationalproblems; in essence, they use DNA to do math. Scien-tists have shown that 1,000 DNA molecules can solve inour months computational problems that would requirea century or a computer to solve.

Other biological molecules are assisting in our continualquest to store and transmit more inormation in smallerplaces. For example, some researchers are using light-ab-sorbing molecules, such as those ound in our retinas, toincrease the storage capacity o CDs a thousand-old.

Some applications o nanobiotechnology include

n increasing the speed and power o disease diagnostics.

n creating bio-nanostructures or getting unctionalmolecules into cells.

n improving the specicity and timing o drug delivery.

n miniaturizing biosensors by integrating the biologi-cal and electronic components into a single, minutecomponent.

n encouraging the development o green manuacturingpractices.

Microarrays

M icroarray technology is transorming laboratoryresearch because it allows us to analyze tens othousands o samples simultaneously.

Researchers currently use microarray technology tostudy gene structure and unction. Thousands o DNA orprotein molecules are arrayed on glass slides to create

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DNA chips and protein chips, respectively. Recent devel-opments in microarray technology use customized beadsin place o glass slides.

DNA MICROARRAYS

DNA microarrays are used to

n detect mutations in disease-related genes.

n monitor gene activity.

n diagnose inectious diseases and identiy the best anti-biotic treatment.

n identiy genes important to crop productivity.

n improve screening or microbes used in bioremediation.

DNA-based arrays will be essential or converting the rawgenetic data provided by the Human Genome Project andother genome projects into useul products. Gene sequenceand mapping data mean little until we determine what thosegenes dowhich is where protein arrays come in.

PROTEIN MICROARRAYS

While going rom DNA arrays to protein arrays is a logi-cal step, it is by no means simple to accomplish. Thestructures and unctions o proteins are much morecomplicated than that o DNA, and proteins are less

stable than DNA. Each cell type contains thousands odierent proteins, some o which are unique to thatcells job. In addition, a cells protein prole varies withits health, age, and current and past environmentalconditions.

Protein microarrays will be used to

n discover protein biomarkers that indicate disease stages.

n assess potential ecacy and toxicity o drugs beoreclinical trials.

n measure dierential protein production across cell

types and developmental stages, and in both healthyand diseased states.

n study the relationship between protein structure andunction.

n assess dierential protein expression in order to iden-tiy new drug leads.

n evaluate binding interactions between proteins andother molecules.

The undamental principle underlying microarray tech-nology has inspired researchers to create many types o

microarrays to answer scientic questions and discovernew products.

TISSUE MICROARRAYS

Tissue microarrays, which allow the analysis o thousandso tissue samples on a single glass slide, are being usedto detect protein proles in healthy and diseased tissuesand validate potential drug targets. Brain tissue samplesarrayed on slides with electrodes allow researchers tomeasure the electrical activity o nerve cells exposed tocertain drugs.

WHOLE-CELL MICROARRAYS

Whole-cell microarrays circumvent the problem o pro-tein stability in protein microarrays and permit a moreaccurate analysis o protein interactions within a cell.

SMALL-MOLECULE MICROARRAYS

Small-molecule microarrays allow pharmaceuticalcompanies to screen ten o thousands o potential drugcandidates simultaneously.

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T

he previous section describes the undamental sci-entic and technological advances which togetherconstitute biotechnology. Here we describe some

o the many tangible rewards aorded by biotech.

Both academic and industrial scientists have come todepend on various biotechnologies to study the workingso biological systems in remarkably precise detail. Thesebiotech research tools have allowed them to answer long-standing scientic questions and have changed the ques-tions they ask, the problems they tackle and the methodsthey use to get answers.

Using the wealth o inormation this research provides,companies then rely on biotechnology tools and tech-niques throughout product development and commer-

cialization.

Research Applications OfBiotechnology

Researchers use biotechnology to gain insight intothe precise details o cell processes: the specic tasksassigned to various cell types; the mechanics o cell divi-sion; the fow o materials in and out o cells; the path by

which an undierentiated cell becomes specialized; and

the methods cells use to communicate with each other,coordinate their activities and respond to environmentalchanges.

Researchers dissect these processes into the smallestpossible bits o useul inormation. This requires iden-tiying the molecular players involved in each acet othe process, elucidating the nature o their interactionsand discovering the molecular control mechanismsthat govern these interactions. Once they have teasedapart details o the process, they must then reassemblethe pieces in a way that provides insight into the inner

workings o cells and, ultimately, o whole organisms.

Interestingly, the tools o biotechnology have also becomeimportant research tools in many branches o scienceother than cell and molecular biology, such as chemistry,engineering, materials science, ecology, evolution andcomputer science. The biotech-driven discoveries in theseelds help the biotech industry and others discover anddevelop products, as well as help industries improve theirperormance in areas such as environmental stewardshipand workplace saety.

UNDERSTANDING CELL PROCESSES

Researchers are making considerable progress in chartingthe path o a cell rom a single, ertilized egg to a whole

organism, a eat that has eluded them or decades. Thedevelopment o a multicelled organism rom a single cellinvolves cell prolieration and cell dierentiationgroupso cells becoming specialized, or dierentiated, to perormspecic tasks. Cell dierentiation is the process o turn-ing o certain genes within a group o cells while turningon others. Scientists are optimistic about elucidating themany steps in the dierentiation pathway and identiyingthe external and internal actors regulating the process.The breakthroughs that gave birth to this optimism are thedevelopment o a protocol or maintaining human stemcells in culture and the birth o the cloned sheep Dolly.

A delicate balance exists between factors thatstimulate cell division and those that inhibit it. Anydisruption of this balance leads to uncontrolled cellproliferationcanceror cell death.

For decades we have known the basic requirementsor keeping small numbers o plant and animal cells inculture or many decades. We maintained these culturesprimarily to collect products that cells produce naturally.For example, plant cell culture gives us favors, colors,thickeners and emulsiers or ood processing.

Researchers now are keeping cells in culture to investi-gate the molecular basis o many cell processes, especiallycell growth, prolieration, dierentiation and death.

All cells progress through essentially the same cycle: Theyincrease in size up to a certain point, the genetic materialreplicates, and the cell divides in two. Understanding whatcontrols the cell cycle is essential to understanding thecause o many human and animal diseases, the basis oincreasing crop plant yields, and a means or quickly in-creasing the cells used to manuacture products as diverseas ermented oods and medicines.

Improvements in cell culture technology have allowed usto better understand the molecular basis o the cell cycle.The rigorously controlled sequence o steps in the cell cycledepends on both genetic and nutritional actors. A delicatebalance exists between actors that stimulate cell divisionand those that inhibit it. Any disruption o this balance leadsto uncontrolled cell prolierationcanceror cell death.

Studying cells in culture has led to a radical revisiono our view o cell death. Formerly we assumed all cells

Biotech Tools in Research and Development

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died through an unorganized, passive mechanism ascell parts and processes gradually deteriorated. Now

we know that much cell death is a highly organized,

well-planned sequence o events programmed into thegenome. Prolonged cell stress and other actors trig-ger programmed cell death, orapoptosis, in which thecell dismantles itsel in an orderly way, breaks downits genome and sends a signal to the immune sys-tem to dispatch white blood cells that will remove it.Programmed cell death eliminates cells with damagedDNA, removes immune system cells that attack healthycells and shapes tissue ormation during development.

A better understanding o cell death can also help usgure out why only some cells with environmentallydamaged DNA turn cancerous; what breaks down in

autoimmune diseases; and how to create better tissuesor replacement therapies.

STEM CELL TECHNOLOGY

Ater animal cells dierentiate into tissues and organs,some tissues retain a group o undierentiated cells toreplace that tissues damaged cells or replenish its supplyo certain cells, such as red and white blood cells. Whenneeded, theseadult stem cells (ASCs) divide in two. Onecell dierentiates into the cell

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