Affinity purification of biologically active and inactive forms of recombinant human protein C produced in porcine mammary gland

Journal of Molecular Recognition : JMR
K E Van CottW N Drohan

Abstract

Recombinant human protein C (rhPC) secreted in the milk of transgenic pigs was studied. Transgenes having different regulatory elements of the murine milk protein, whey acidic protein, were used with cDNA and genomic human protein C (hPC) DNA sequences to obtain lower and higher expressing animals. The cDNA pigs had a range of expression of about 0.1-0.5 g/l milk. Two different genomic hPC pig lines have expressed 0.3 and 1-2 g/l, respectively. The rhPC was first purified at yields greater than 60 per cent using a monoclonal antibody (mAb) to the activation site on the heavy chain of hPC. Subsequent immunopurification with a calcium-dependent mAb directed to the gamma-carboxyglutamic acid domain of the light chain of hPC was used to fractionate a population having a higher specific anticoagulant activity in vitro. The higher percentages of Ca(2+)-dependent conformers isolated from the total rhPC by immunopurification correlated well with higher specific activity and lower expression. A rate limitation in gamma-carboxylation of rhPC was clearly identified for the higher expressing animals. Thus, transgenic animals with high expression levels of complex recombinant proteins produced a lower percentage of biologically active protein.

References

Oct 13, 1992·Annals of the New York Academy of Sciences·W H VelanderJ L Johnson
Aug 1, 1992·Journal of Cellular Biochemistry·T D Wilkins, W Velander
Dec 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·W H VelanderW N Drohan
Mar 13, 1987·Science·C T Esmon
Oct 1, 1988·Thrombosis Research·M J HeebJ H Griffin
Aug 1, 1986·Thrombosis Research·J L Martinoli, K Stocker
Nov 7, 1995·Proceedings of the National Academy of Sciences of the United States of America·R DrewsH Luboń
Nov 1, 1994·Transgenic Research·W N DrohanH Lubon
May 2, 1994·Annals of the New York Academy of Sciences·T MorcölW H Velander
Sep 1, 1993·Protein Science : a Publication of the Protein Society·R M MestersJ H Griffin
May 1, 1992·Biotechnology and Bioengineering·K KangC L Orthner

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Citations

Apr 29, 1998·Journal of Chromatography. a·A DegenerW H Velander
Nov 6, 2001·Journal of Biochemical and Biophysical Methods·T Burnouf, M Radosevich
Nov 13, 2004·The Journal of Nutritional Biochemistry·P A PrietoB Kelder
Jul 5, 2005·Expert Opinion on Investigational Drugs·K E Van Cott, W H Velander
May 7, 2009·Annals of the New York Academy of Sciences·Amy-Lynn FrankshunCarol A Bagnell
Apr 20, 1999·Nature Biotechnology·D C JohnN J Bulleid
Dec 24, 2010·Reproduction : the Official Journal of the Society for the Study of Fertility·Amy-Lynn FrankshunCarol A Bagnell
May 30, 2021·Proceedings of the National Academy of Sciences of the United States of America·Luhan YangGeoffrey Yang

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