00048 j silva proteo monitor 2005 1104

11/4/2005 | Volume: 5 | number 42

in this issue

FeaturesWaters Develops New Label-Free, Mass Spec-Based Method for Absolute QuantificationThe new method relies on an "unexpected" relationship between the average MS signal response for the three most intense trypticpeptides and protein concentration. However, so far the method can only be performed using the Waters Q-TOF Premier instrument,because the tool has a special data-collection strategy that toggles back and forth between acquisition of low-energy precursor ioninformation and elevated energy fragmentation data.

EMBL Researchers Develop High-ThroughputMethod for Producing Monoclonal AntibodiesThe method was able to generate 68 monoclonal antibodies within six weeks after immunizing eight mice with 10 antigens each.

Proteomics PioneerNIH's Glen Hortin on Translating New Biomarkers into Clinical Laboratory Tests

Industry BriefsCaprion, AstraZeneca, Beckman Coulter, Consorta, Beckman Coulter, PerkinElmer, EU, Compugen

New Products/Movers & ShakersInternational School of Advanced BioMedicine and Bioinformatics, Lipari International School for ComputerScience Researchers, Sebastian Meyer-Plath, Jizhong Zhou

ChartsRecent Proteomics Papers of Note (K-Z)*

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Features

Waters Develops New Label-Free, Mass Spec-Based Method for Absolute QuantificationWaters has developed a new label-free, mass spectrometer-based method for absolute quantification ofproteins.

The new method, published in the Oct. 11 issue of Molecular & Cellular Proteomics, relies on an"unexpected" relationship between protein concentration and the average MS signal response for the threemost intense tryptic peptides, according to Jeff Silva, the first author of the study who is a senior researchand development scientist in Waters' Core Technologies/Proteomics department.

There is one important caveat to the method, Silva noted: So far, the method can only be performed usingthe Waters Q-TOF Premier mass spec, because the instrument has a special data-collection strategy thattoggles back and forth between acquisition of low-energy precursor ion information and elevated energyfragmentation data.

Normally, the relative quantitation of a peptide as compared to another peptide can not be determined bycomparing MS intensities because peptides ionize at different efficiencies, Silva explained. The newabsolute quantification method is based upon a relationship that was discovered between absoluteconcentrations of proteins and the [MS signal] "response factor" of the three highest ionizing peptides of agiven protein.

"It turns out that at equimolar concentration, the average intensity measurement of those top ionizingpeptides are the same," said Silva. "So there's a counts-per-mole equivalent for any given protein."

Silva's research team tested the method of absolute quantification byfirst working with five proteins of known concentrations that varied"It seems simple. They've in size from 15 to 98 kilodaltons, then carrying out the same

noticed a trend where if you calculations with a subset of human serum proteins of unknownlook at the most abundant concentrations.ions, you can generate a

The researchers calculated the average signal response from the topuniversal factor that you canthree tryptic peptides for each of the proteins, which varied in

multiply everything with. You concentration from six picomoles to 15 pmoles. They showed thatfind your scaling factor, and each of those proteins had the same response factor, such that theyou can go back and actually signal response is a fixed number of counts per picomole of protein.

figure out micromolarOnce they had the response factor for their Waters Q-TOF Premieramounts." instrument, the researchers used that factor to calculate the absolutequantity of 11 proteins in human serum. The concentrations of theproteins as determined by the new method correlated with

concentrations of the proteins that had been previously published in other studies.

"It seems simple," said Minerva Hughes, a recent PhD graduate in Craig Townsend's laboratory at JohnsHopkins University who has been using the Waters Protein Expression system to do relative proteinquantitation. "They've noticed a trend where if you look at the most abundant ions, you can generate auniversal factor that you can multiply everything with. You find your scaling factor, and you can go backand actually figure out micromolar amounts."

The reason that only Waters Q-TOF Premier instruments work with this method is that so far, only thoseinstruments collect data in an alternating fashion, such that there are three data points — mass, retention


time, and signal intensity — per time interval, Silva said. Other instruments acquire data through only afraction of the peak list, until they have gathered enough information to identify that peptide.

"The alternate scanning mode of acquisition allow you to get a true sense of what's there in terms of peakarea to a given protein," said Silva. "Because we were able to see the true 3D peptide mass fingerprint,without partial peak sampling, the correlations fell right out of the data."

Silva and Hughes noted that absolute quantitation is important for determining the stoichiometry ofmolecules that make up a complex.

Paul Skipp, the manager of the Center of Proteomics Research at theUniversity of Sussex in the U.K., added that absolute quantitation is"I think the way that [mass important for understanding signaling pathways.

spec] technology is going tomove in the future is to move "In a signaling pathway, you tend to have a small signal that starts

that causes a protein to bring other proteins to it — that may requireaway from labeling."10 molecules, and those 10 molecules may signal to another protein,"he said. "If you know a cell in its normal state has so many molecules

of Y and so many molecules of Z, you start to understand how that cell's working. Also, if one of thosemolecules starts to produce a larger number of molecules, that could be an indicator of an onset of disease."

13Skipp currently uses a technique called AQUA to do absolute quantitation. The method relies on using aC-labeled molecule as an internal standard. Skipp had not heard of Waters' new method of absolutequantitation, which does not require protein labeling, but said that he was pleased with the way Waters'Protein Expression system had worked for relative quantitation.

"I think the way that [mass spec] technology is going to move in the future is to move away from labeling,"said Skipp. "The beauty of the Protein Expression is that you don't have to label, and you can compare morethan four conditions, and you can compare different experiments to each other."

Silva said Waters is currently integrating the new absolute quantification relationships into their software sothat it will be easy to calculate the average intensities of the top three ionizing peptides. He noted that thereason for using the top three ionizing peptides, as opposed to another number, was a function of the size ofthe protein. For small proteins, the top two might work better, and for large proteins the top four might workbetter, he said.

"You could make it a function of molecular weight, but three worked in this demonstration," he said.

— Tien Shun Lee ([email protected])


EMBL Researchers Develop High-ThroughputMethod for Producing Monoclonal AntibodiesUsing antigen arrays, researchers at the European Molecular Biology Laboratory have developed ahigh-throughput method for producing monoclonal antibodies.

In a study published in the Oct. 27 issue of Proteomics, researchers led by Federico De Masi and AlanSawyer showed that the method was able to generate 68 monoclonal antibodies within six weeks afterimmunizing eight mice with 10 antigens each.

"Normal antibody production takes five to nine months for a single antibody, depending on howimmunogenic the antigen is," said De Masi, who is now a postdoc in Martha Bulyk's laboratory at Brigham& Women's Hospital. "By using antigen microarrays, we can basically detect the antibody of the animal at amuch earlier stage, and the screening is faster than with an ELISA or Western blot."

The EMBL's patented antibody production method has been commercialized by Taiwan-based Abnova,which signed a collaboration agreement with EMBL in December 2004.

As with many other antibody production methods, the EMBL method begins with immunizing a mouse withan antigen of interest. Researchers then wait four weeks for the mouse to generate an immune responsebefore sacrificing it.

After B-cells have been harvested from the spleen of the mouse, researchers generate a library of thousandsof hybridoma cell lines — immortal cell lines that produce one type of monoclonal antibody each — byfusing the B cells with cells from a mouse cancer cell line.

The key to the new antibody production method is that screening ofhybridoma cell lines is done using a chemically modified glass slide"The greatest challenge in coated with the antigen of interest. Because this array method is more

creating the array is that we sensitive for detecting antibodies than ELISA or Western blothad to find the right methods, researchers do not have to wait as long for mice to generate

an antibody response before sacrificing the animals.concentration of the antigens,and the right chemical

While traditional monoclonal antibody production methods requiremodification of the slide to waiting six weeks to three months before sacrificing the immunizedget the microarray functional. mouse, the EMBL method requires only four weeks of waiting.Then each sample that we

After hybridomas have been spotted onto the antigen-coated slide, thespot has a different densityslide is washed, incubated with a fluorescent secondary antibody thatand viscosity, so there was a recognizes mouse antibodies, then washed again. If an antibody has

lot of tweaking that had to be stuck to the antigen-coated slide, then it can be visualized through thedone. It took some time to fluorescent secondary antibody.find the right conditions that

"The greatest challenge in creating the array is that we had to find thewere highly reproducible forright concentration of the antigens, and the right chemical

the whole spectrum of modification of the slide to get the microarray functional," said Deantibodies." Masi. "Then each sample that we spot has a different density and

viscosity, so there was a lot of tweaking that had to be done. It tooksome time to find the right conditions that were highly reproducible

for the whole spectrum of antibodies."

To make the monoclonal antibody production process even more high throughput, EMBL researchersdecided to immunize mice with more than one antigen at a time. They first tried two antigens per mouse,


then five, then 10.

In a large-scale experiment, De Masi and his colleagues immunized eight mice with 10 antigens each. Afterthe mice were sacrificed and hybridomas were created, the researchers screened the hybridomas on 10 slidesthat were coated with one antigen each. In the end, the researchers were able to generate antibodies against68, or 85 percent, of the antigen targets, within six weeks of primary immunization.

Aside from using the arrays to cut down on antibody production time, EMBL researchers also saved on timeby using custom-built robots to automate the hybridoma production process.

"The robots take care of changing cell medium, fusion of B-cells, plating onto plates, [and] incubation," saidDe Masi. "Otherwise, we'd have to have somebody 24 hours per day taking care of these cells."

Currently, the EMBL's monoclonal antibody core facility will produce monoclonal antibodies to an antigen!for about 2,500 ($3,017) per antigen, De Masi said. Monoclonal antibodies against peptides cost a little €more — 3,000 ($3,591) per peptide — because it is harder to raise a good immune response against a

peptide, which is generally shorter than an antigen.

The EMBL generally leaves quality testing of antibodies up to the researchers who have requested them, DeMasi said.

"Generally what's done is we obtain five to 10 strong affinity hybridomas and we send it to the investigatorso that he or she can choose the one that he or she likes," said De Masi. "Depending on what the investigatorneeds the antibody for, the antibody might be fantastic for one application, but useless for another. Forexample, with some of our antibodies that we did test, they gave some signal on the microarray, a beautifulWestern blot band, but no signal on an ELISA."

Abnova president and CEO Wilber Huang said that his company's collaboration with EMBL furthers itseffort to become a leader in high-throughput antibody production.

"Custom service and partnership now form the core of our business. We are concurrently developing acomprehensive antibody bio-tool catalog for the drug discovery and diagnostic industry," said Huang,following the formation of the collaboration agreement. "As such, we are focusing on several key areas ofhuman proteins: kinase, apoptosis, receptors, stem cell and plasma proteome. The collaboration with EMBLis the latest step in our effort to realize these goals."

— Tien-Shun Lee ([email protected])


Proteomics Pioneer

NIH's Glen Hortin on Translating New Biomarkers into Clinical LaboratoryTests

At A Glance

Name: Glen Hortin

Position: Chief of clinical chemistry, Department of Laboratory Medicine at theNational Institutes of Health, since 2000. Acting, assistant chief of clinical chemistry,since 1997.

Background: Associate professor of pathology, University of Alabama atBirmingham's clinical chemistry section, 1993-1997.

Glen Hortin Assistant professor in pediatrics and pathology, Washington University, 1989-1992.Chief of clinical

chemistry,Department of

Laboratory MedicineLast week, Glen Hortin gave a talk on "Translating New Biomarkers into ClinicalNational Institutes of

Health Laboratory Tests" during an American Association for Clinical Chemistry meeting inWashington, DC. ProteoMonitor caught up with Hortin to find out about his advice

on this issue, and about his research background.

How did you get into developing biomarkers and clinical tests?

Originally, I was interested mostly in protein chemistry. I started in the 1970s looking at proteinbiosynthesis. As an undergraduate and graduate student, I was looking at processing events like theprocessing of secretory proteins and glycosylation of proteins. As a graduate student I worked on the basicaspects of a signal hypothesis in terms of how secretory proteins are processed.

Then I was interested generally in the issues of protein processing — post-translational modifications andmolecular recognition between proteins, how various proteases or substrates would find their right substratesor protease to inhibit — how they got together. Simultaneously I was getting interested in clinical chemistry.Those are interests of mine that go back about 20 years.

Which laboratory were you working in?

I was working with Irving Boime at Washington University in St. Louis. I continued on as a faculty person.Arnie Strauss helped me get going — he had done a lot of work in the protein processing area. He was apediatric cardiologist, but also did basic science.

Before I got started on my graduate work also, I had the good fortune to work with Donald Steiner at theUniversity of Chicago — he was the fellow who discovered proinsulin. At the time, he was studying theprecursors of proinsulin. I spent a couple of summers in his lab as a student. It was an interesting entré intoprotein chemistry.

I went to medical school, and I did a residency from 1983 to 1987 in clinical pathology. My main interestwas in the clinical chemistry end of the laboratory.

What types of problems were you working on?


In clinical chemistry, you tend to get interested in a wide range of problems — whatever problems come upin the laboratory. It's really kind of problem solving and problem focused. In the clinical chemistry area,probably half of the assays we deal with relate to protein measurement — some of them are enzyme assays,some of them are immunoassays, some of them are simple electrophoretic assays. Basically, in the clinicallaboratory, a major portion of the protein assays are really kind of within the subspecialty of clinicalchemistry. Another section is the immunology section.

Were you working on enzyme assays?

Well, I did a fair amount of work in looking at protease specificity. From a practical standpoint, you're oftenin a problem solving mode in the clinical lab to address the patient needs. In general, I've had a more basicresearch lab effort that's kind of gone in parallel. Part of that effort was directed at studying post-translationalprocessing of proteins. I spent a fair amount of work on sulfation of proteins.

Where did you go after Washington University?

I stayed on as faculty there for about five years, and then I went to the University of Alabama inBirmingham. A greater amount of my time there was spent doing clinical work — helping to run a clinicallaboratory. I got involved fairly actively in terms of point-of-care testing, which is a lot of the testing done atthe patient's bedside.

The things done at the bedside are usually things like glucose testing, electrolytes, things like that.

Then eight years ago, I came to the NIH, where I had a little bit more time for doing research work. I reallybecame interested in some of the issues with protein analysis. With new developments in mass spectrometryand new tools, I could go back and look at some of the issues I had been interested in years before. I supposeI was basically interested in proteomics 15 or 20 years ago, but we didn't have the tools to do it. I remember15 years or so ago being interested in 2D electrophoresis and how you might use it for diagnostic purposes.But it wasn't well enough developed then. You would get lots of spots, but you really couldn't identify whatmost of the spots were.

It's really over about the last five years or so that with new mass spectrometry tools, and with completion ofthe human genome that basically defines all the protein sequences, you can go back and identify whateverprotein or peptide you want, and you have a lot more of a solid basis for doing these things. Another thing isthat detection limits have become much more practical to work with. The scale of samples that you wouldget in a clinical lab before — you might have had to draw much larger volumes of blood than would beroutinely practical, and kind of go through many steps to get down to what you need. This is starting to reallyenable things to be more approachable on true clinical samples.

Did you start to work with biomarkers at the NIH?

Well, 'biomarkers' is in a sense a nonsense term. I've been working with biomarkers for 30 years, thoughpeople didn't call them that back then. All the lab tests that we deal with pretty much are measuringbiomarkers. Biomarkers is such a vague term, it's practically a nonsense term. I don't know why people useit. It's just kind of a catchy term that for some reason people think means something special.

Every test that we use in the clinical lab is basically measuring a biomarker — it may be an enzyme activity.If you look at the current definition, it's basically anything that you can currently measure that might berelated to a physiological response that you might be interested in following. We've had biomarker labs formany years — they're called clinical labs.

It is a very interesting time in that you can sort through possibilities much faster in terms of doing discovery


work, and the whole pace of discovery has been speeded up a lot. Some of the techniques allow you to lookat hundreds of components at a time. That's really a new dimension in terms of looking at things. In the past,most of the things that we've done in the clinical lab have been done one test at a time. That's not the case100 percent [of the time], but the majority of them have been. A few things we've looked at [involved morethan one thing at a time] — a profile on electrophoresis, or some things looked at an entire pathway, orperhaps 10 or more components. But we now have a new tool set to look at these things, and I think it isopening up new possibilities, particularly to look at molecular variation and some of the post-translationalmodifications.

What kind of tests are you working on developing currently?

Mainly I'm interested at the moment in trying to characterize a diversity of smaller peptide components inbiological fluids — mainly plasma and urine samples. By traditional tools, that was one area that we did notsee in very great detail. The new tool sets have really expanded quite a bit our ability to look at these. Massspec is very good for looking at peptides. The older tools like 2D electrophoresis — the peptides fall off thebottom, so they're kind of invisible. We probably had underaccounted for them some. So that's the area I'minterested in looking at the moment.

What issues do researchers have to deal with in translating a biomarker into a clinical test?

There are many steps in going from a basic biomarker to a useful laboratory test. Particularly in some of theprofiling techniques that have been used for the MALDI-TOF or SELDI-TOF techniques have not appliedthe usual types of standards that we use in the clinical laboratory. To start to figure out how to standardizethese types of assays better — you have to define some of the things that you usually do for any clinical testin terms of evaluating reproducibility, linearity of responses. You have to figure out how the intensity ofvarious peaks relate to the concentration of what you're trying to measure. I think that to move some of thesetechniques to clinical lab tests, you have to come up with approaches that deal with these things. Otherwiseyou would not be meeting the usual standards that we would apply in a clinical lab.

What's the biggest challenge in moving something into a clinical test?

There are so many steps in this that it's a little bit hard to know where to begin. There are really probably ahalf dozen or more important steps. There are whole issues in terms of having adequate specimens tovalidate the test in the first place, and to kind of serve as a database. There are so many issues related to thespecimen selection, and how you perform those studies. And then people are still trying to grapple withspecimen collection issues. Often times they haven't paid quite enough attention in terms of pre-analyticalvariables — the patient preparation, and how to standardize the collection processes and things.

There's also the issue of marker selection. Many markers are not going to be suitable if they're too variable orunstable.

Most of what we're doing at the moment is method and technology development rather than specific testdevelopment. It's a little premature to try to develop a test until you really have your methods optimizedmore. A lot of our efforts now are really towards trying to get the most information possible about peptiderepertoires in different samples and stability issues so that we can figure out how to handle the samplesbefore we even get to the point of doing a test. Once we do that, I think then we'll be able to move forwardfairly quickly and a little bit more rigorously in terms of doing tests.

We have analyzed some clinical sets and things, but those were a little bit more discovery efforts in terms offinding peptides that might be discriminatory for certain things. We weren't really to the point of doing testsyet.

Some people think as soon as they get some peaks showing up in a pattern, they have a test, but I think in


general there's a fairly big gap in terms of finding a few peaks in a pattern and figuring out how you can usethat meaningfully in a test.

What kind of advice would you give to someone who has found some biomarkers and is looking todevelop them into a clinical test?

Generally, I feel it's important to be able to identify what your peaks are, and to understand the biology andwhat they represent. Is there really any physiological basis that they should be related to what you're trying tounderstand? The second thing is you have to optimize your methods so you get good reproducibility. Yourability to get accurate measurements in general kind of depends on your ability to get high signal to noise. Ifyou're dealing with very weak signals, in general you're not going to be able to get very robust tests out of it.


Industry Briefs

Caprion Partners with AstraZeneca on Prostate Cancer Targets

Caprion and AstraZeneca will collaborate to develop therapies for prostate cancer, Caprion Pharmaceuticalssaid this week.

Under the agreement, AstraZeneca will evaluate prostate cancer drug targets discovered by Caprion andobtain exclusive, worldwide rights to develop and commercialize therapeutic applications for selectedtargets. Caprion will retain rights to other targets.

AstraZeneca will pay Caprion an unspecified amount for up-front payment and license fees. The companywill make additional payments contingent on its meeting certain development and commercializationmilestones.

Beckman Coulter Licenses Clinical Lab Tests to Consorta; Deals Worth $105M Over Five Years

Beckman Coulter will provide Consorta its chemistry, immunoassay, hematology, and automation productlines, the company said this week.

The three contracts are worth approximately $105 million over the next five years to Beckman. Consortaalso extended its hematology agreement with Beckman, valued at $10 million a year, for two more years,Beckman said.

The contract provisions include the company's new line of UniCel systems, such as the DxC 600 and 800chemistry analyzers, the DxI 800 Access immunoassay system, and the Power Processor automation system,said Robert Kleinert, an executive vice-president at Beckman Coulter, in a statement.

Consorta will also have access to nearly 200 different reagent kits.

Beckman Coulter Q3 Revenues Grow 2 Percent as Income Drops Nearly 40 Percent

Beckman Coulter this week said that its third-quarter sales grew by a 2 percent, while net income fell 38percent.

Total worldwide sales for the three months ended Sept. 30 inched up to $593.4 million from $581.2 millionin the same quarter last year. The company reported that $591.7 million of total sales for the quarter wereorganic, while $1.7 million could be attributed to currency.

The Fullerton, Calif.-based company also said that sales in the US remained even compared to last year, butthat international sales grew by 3.8 percent.

Net income in the third quarter fell to $36.2 million from $57.2 million year over year, the company said,resulting in basing earnings being reduced to $0.58 per share from $0.93 per share.

Beckman Coulter said that the company incurred $19.2 million in special charges during the quarter, mostlydue to "impaired receivables and leased equipment" related to Hurricane Katrina — which cost the company


$4.9 million — and "the non-cash write off of intangible assets of discontinued robotic automation product"from the discontinuation of the Bovine Spongiform Encephalopathy testing initiative, which cost Beckman$13.4 million.

The company added specifics to a plan to reorganize the company, as reported by ProteoMonitor's sisterpublication GenomeWeb News on July 22, signaling its intent to "close at least three facilities, sell twoparcels of real estate, harvest or discontinue mature product lines," reduce 350 positions, and streamline thecompany's physical distribution.

Beckman Coulter president and CEO Scott Garrett said in a statement that he expects the lay-offs will resultin a benefit of up to $2 million in the fourth quarter of this year, as well as "$15 million of annual savings in2006, growing to $20 million of annual savings in 2007 and beyond."

Garrett also offered a preliminary outlook on 2006. Beckman is now expecting total 2006 sales to be$2.5-$2.6 billion. He added that the company's decision to favor operating-type leases over sales-type leasesfor instrument placements may reduce sales over 2005 and 2006 as the change in policy is implemented.

"Our original range for the impact of the leasing policy change on sales through 2006 was $200 to $220million," Garrett said. "We now expect the change to reduce revenues by $190 to 200 million over theimplementation period, which will be split about evenly between 2005 and 2006."

Beckman Coulter spent $51.6 million on R&D in the quarter, compared to $51.5 million in the same quarterlast year.

As of Sept. 30, the company held cash, cash equivalents, short-term investments, and restricted cash totaling$59.3 million.

PerkinElmer Obtains $350M Revolving Loan; Cash Could Fund M&A, Alliances

PerkinElmer obtained a $350-million unsecured revolving credit facility, the company said this week.

The five-year loan, which replaces a previous $100 million facility, will be used for "general corporatepurposes," such as working capital, refinancing, capital expenditures, share repurchases, acquisitions, andstrategic alliance, said PerkinElmer.

The facility was jointly made with Banc of America Securities and Citigroup Global Markets.

EU Awards $10.8M to Form Network of Excellence on Biological Research

€The Commission of the European Union awarded 9 million ($10.8 million) to establish a virtual institute for biological research, the European Bioinformatics Institute said this week.

ENFIN, or the Experimental Network for Functional INtegration, will combine 20 computational andexperimental biology labs across 17 institutions in 13 counties to make computational systems biologyaccessible to European scientists. While applicable to any area of biological research, the network will focuson the regulation of cell division.

While there's an open-access database for almost every type of biological information, the average biologiststruggles to access the data, the institute said.


"Researchers will be able to go straight to the public data that they want, combine it with their ownunpublished data and perform truly integrated analyses using data from different types of experiments," saidthe institute's Ewan Birney, who will coordinate ENFIN.

ENFIN will incorporate both "wet" and "dry" biologists whose expertise spans database architecture, dataanalysis tools, and experimental molecular biology.

Compugen Reports 22-Percent Slide in Q3 Revenue, Names New CEO

Compugen said this week that revenues for the third quarter declined 22 percent while net losses remainedflat.

The company also said it has named Alex Kotzer president and CEO. He started on Sept. 1.

Total revenues for the three months ended Sept. 30 fell to $761,000 from $971,000 year over year. About87.5 percent of the amount, or $666,000, came from "governmental and other grants," said Compugen

Third quarter R&D spending increased slightly to $3 million from $2.8 million in the previous year.

The company reported that net loss remained flat for the quarter, at approximately $3.6 million, or $.13 pershare.

As of Sept. 30, the company had $29.9 million in cash and cash equivalents.


New Products/Movers & Shakers

New Products

The International School of Advanced BioMedicine and Bioinformatics and the Lipari InternationalSchool for Computer Science Researchers are jointly organizing a two-week course on "Proteomes andProteins." The course will be held on Lipari — an island off the coast of Eastern Sicily — from July 9 toJuly 22, 2006. It will focus on describing knowledge of proteomes, their dynamic changes during the lifecycle of cells and organisms, and their modifications in disease states.

More information on the course is available at http://lipari.cs.unict.it/lipari/currentedition.htm.

Movers & Shakers

Sebastian Meyer-Plath returned to Bruker Daltonics as vice president for its nuclear, biological, andchemical business and as a managing director of Bruker Daltonik GmbH, the Bruker Daltonik operation inLeipzig, Germany, the company said this week.

He originally joined Bruker in 1993, and held several project- and product-management positions in thecompany. He left in 2002 for Germany's Advalytix AG and joined Smiths Detection as vice president ofsales & marketing in 2003.

At its Oct. 25 meeting, University of Oklahoma's Board of Regents decided to bring Jizhong Zhou to theuniversity as a functional genomics researcher, said Lee Williams, vice president of research. Zhou'sgenomics laboratory on campus is expected to be fully operational by January 2006.

Prior to his OU appointment, Zhou was in Oak Ridge, Tenn. at Oak Ridge National Laboratory.


Charts

Recent Proteomics Papers of Note (K-Z)*

Journal Title Authors

Molecular & CellularAbsolute quantification ofProteomics. Silva JC, Gorenstein MV, Li GZ, Vissers JP,proteins by LCMSE: A virtue2005 Oct 11; Geromanos SJ.of parallel MS acquisition.[Epub ahead of print]

Molecular & Cellular Biomarker discovery from Gronborg M, Kristiansen TZ, Iwahori A,Proteomics. pancreatic cancer secretome Chang R, Reddy R, Sato N, Molina H,2005 Oct 8; using a differential proteomics Jensen ON, Hruban RH, Goggins MG, Maitra[Epub ahead of print] approach. A, Pandey A.

High throughput quantitativeMolecular & Cellular glycomics and Uematsu R, Furukawa JI, Nakagawa H,Proteomics. glycoform-focused proteomics Shinohara Y, Deguchi K, Monde K,2005 Sep 16; of murine dermis and Nishimura SI[Epub ahead of print] epidermis.

Molecular & Cellular Comparative proteomicsProteomics. analysis of intra- and Hu Y, Malone JP, Fagan AM, Townsend RR,2005 Sep 30; inter-individual variation in Holtzman DM.[Epub ahead of print] human cerebrospinal fluid.

Differential protein expressionMolecular Psychiatry. in the prefrontal white matter Alexander-Kaufman K, James G, Sheedy D,2005 Sep 20; of human alcoholics: a Harper C, Matsumoto I.[Epub ahead of print] proteomics study.

Liver tumors in wild flatfish: a Stentiford GD, Viant MR, Ward DG, JohnsonOMICS. histopathological, proteomic, PJ, Martin A, Wenbin W, Cooper HJ, Lyons2005 Fall;9(3):281-99. and metabolomic study. BP, Feist SW.

Proceedings. BiologicalBehavioural manipulation in aSciences/The Royalgrasshopper harbouring Biron DG, Marche L, Ponton F, Loxdale HD,Society.hairworm: a proteomics Galeotti N, Renault L, Joly C, Thomas F.2005 Octapproach.22;272(1577):2117-26.

Proceedings of theNational Academy of

An evolutionary proteomicsSciences of the Unitedapproach identifies substrates Budovskaya YV, Stephan JS, Deminoff SJ,States of America.of the cAMP-dependent Herman PK.2005 Sepprotein kinase.27;102(39):13933-8. Epub

2005 Sep 19.

Proteomics. Proteomics-based consensus2005 Oct 11; prediction of protein retention Tjalsma H, van Dijl JM.[Epub ahead of print] in a bacterial membrane.

Functional proteomics andcorrelated signaling pathway

Proteomics. of the thermophilic bacterium Topanurak S, Sinchaikul S, Sookkheo B,2005 Oct 13; Bacillus stearothermophilus Phutrakul S, Chen ST.[Epub ahead of print] TLS33 under cold-shock


stress.

Proteomics.Proteomic dataset of mouse Mayr U, Mayr M, Yin X, Begum S, Tarelli E,2005 Oct 20;aortic smooth muscle cells. Wait R, Xu Q.[Epub ahead of print]

Proteomics.Proteomic dataset of Sca-1(+) Yin X, Mayr M, Xiao Q, Mayr U, Tarelli E,2005 Oct 20;progenitor cells. Wait R, Wang W, Xu Q.[Epub ahead of print]

Printing of proteinProteomics. microarrays via a Barron JA, Young HD, Dlott DD, Darfler MM,2005 Sep 30; capillary-free fluid jetting Krizman DB, Ringeisen BR.[Epub ahead of print] mechanism.

Proteomics.A proteomics approach to2005 Sep 30; Wagg SK, Lee LE.identifying fish cell lines.[Epub ahead of print]

A top-down proteomicsProteomics. approach for differentiating Williams TL, Monday SR, Edelson-Mammel2005 Sep 30; thermal resistant strains of S, Buchanan R, Musser SM.[Epub ahead of print] Enterobacter sakazakii.

Proteomics. Large-scale analysis of theMatsumoto M, Hatakeyama S, Oyamada K,2005 Sep 30; human ubiquitin-relatedOda Y, Nishimura T, Nakayama KI.[Epub ahead of print] proteome.

Characterization of mediumconditioned by irradiated cells

Radiation Research. Springer DL, Ahram M, Adkins JN, Kathmannusing proteome-wide,2005 Nov;164(5):651-4. LE, Miller JH.high-throughput mass

spectrometry.

Differential expressionprofiling of membraneproteins by quantitative

Stem Cells. Foster LJ, Zeemann PA, Li C, Mann M,proteomics in a human2005 Oct;23(9):1367-77. Jensen ON, Kassem M.mesenchymal stem cell line

undergoing osteoblastdifferentiation.

*A-J were published in last week's ProteoMonitor


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00048 j silva proteo monitor 2005 1104

Documents

new method

absolute quantification

mass spec based method

ms signal response factor

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new labelfree