blood products production from transgenic animals

Animal cell culture is used to produce variety of commercial & medicinal prods.

Evaluation of new drugs on culture cells is being done which is cost-effective and less time consuming.

Whole blood consists of RBC’s,WBC’s,platelets are suspended in fluid called plasma.

when blood is allowed to clot it forms cross-linked network known as serum.

Various comp of blood serve a wide range of physiological func.

Specific blood prtns that are of therapeutic use are:-

clotting factors Serum albumin.

fibrinolytic agents. Ig.

T-PA is an enzyme produced by mammalian cells in small amts.

This removes blood clot and obstruction in blood vessels.

First prtn to be produced on a commercial scale.

GENETECH had cloned human T-PA for treating blood clots.

The trade name of this product is Activase.

Human mRNA coding for T-PA from human mylenoma cell line.

Reverse transcription converted to cDNA

Inserted to Chinese Hamster Cell lines

CHO cells excrete T-PA in medium

Isolation of T-PA

blood loss must be stopped in case of injury in which blood vessels are damaged.ss

prtn fibrinogen fibrin. aggregation of platelets at site of injury stops blood flow

blood coagulation process is dependent on various clotting factors[factor I-XIII].

factor VIII prevents hemophilia.

In hemophiliac patients blood keeps flowing out in case of injury.

In 1984,gene that controls production of F VIII was isolated.

This enabled research & development of genetically engineered clotting factor.

In 1992, 1st genetically engineered clotting factor was introduced under trade name RECOMBINATE & RE FACTO.

In 1993, GENETECH produced 2nd clotting factor under trade name KOGENATE.

There are 4 other recombinant F VIII:- BIO GEN GENETIC

THERAPY BIO VECTOR IMMUNE

RESPONSE

EPO is a glycoprtn hormone produced primarily by cells of kidney.

Regulates RBC production.Secondary amts of hormone is synthesized in liver of healthy adults.

This sialoglycoprotein hormone consists of 165 amino acids that form a single polypeptide chain containing two intra-chain disulfide bonds (Cys7-161 and Cys29-33) and four potential glycosylation sites, three of which are N-linked (Asn24, Asn38 and Asn83) and one O-linked (Ser126) (1,2).

The molecular mass of EPO is 30-34 kDa, but the molecular mass of the peptide chain is only about 18 kDa. Thus, about 40% of the glycosylated EPO molecule consists of carbohydrate, mostly in the form of N-linked complex-type glycans.

Recombinant human EPO (rhEPO) is produced commercially by the expression of EPO cDNA clones in eukaryotic cell lines, most commonly in Chinese hamster ovary or baby hamster kidney cells with CMV and EF-1 promoters.

Transgenic pig containing the the human EPO gene placed under EF 1 promoter was developed

F2 generation piglets was analysed for pres of rHu-EPO recovery of rHu- EPO two-step precipitation method for the recovery of the

recombinant human erythropoietin (rhEPO) ammonium sulfate and divalent metal ions yield was

82.1%

rHu- EPO is used for variety of clinical situations

increased production of RBCs anemia in HIV patients. curing post operative anemia.

Albumin from plasma fractionation is generally considered to be a safe and effective product.

A number of factors makes recombinant albumin manufacture increasingly attractive. These factors are as follows:

The perceived risk of viral contamination in serum albumin

likelihood of reduced availability of recovered plasma in the future

price

Major component of blood plasmaFluid balance b/w blood & tissuesRegulates transport of Amino acids, Fatty

acids and Hormones, Drugs and drug metabolites

The expression of human albumin in the milk of transgenic cows has been identified as the most promising approach.

A mammary gland expression vector that contained the goat β-casein gene promoter, 5′upstream regulatory region, exons 1,2, intron 1 as well as the human serum albumin (hALB) mini-gene (including the full-long sequences of hALB cDNA and its intron 1).

In exhaustible & in expensive.

Free from infectious agents.

New blood prtns not available earlier .