Transgenic pigs as bioreactors

A comparison of gamma-carboxylation of glutamic acid in recombinant human protein C and factor IX by the mammary gland

Kevin Van Cott, Stephen P. Butler, Christopher G. Russell, Anuradha Subramanian, Henryk Lubon, F. C. Gwazdauskas, James Knight, William N. Drohan, William H Velander

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

The mammary gland of transgenic livestock can be used as a bioreactor for producing complex therapeutic proteins. However, the capacity for making a given post-translational modification upon any given polypeptide is uncertain. For example, the efficiency of gamma-carboxylation of glutamic acid in the amino terminal regions of recombinant human protein C (rhPC) and recombinant human Factor IX (rhFIX) is different at similar expression levels. At an expression level of about 200 μg/ml in the milk of transgenic pigs, rhFIX is highly gamma-carboxylated as indicated by pro-coagulant activity and amino acid sequencing. However, only about 20-35% of rhPC has a native, gamma-carboxyglutamic acid-dependent conformation and anti-coagulant activity. Thus, this work provides an example of apparent differences in substrate specificity between two homologous proteins to the endogenous carboxylase of porcine mammary epithelium which leads to varying degrees of post-translational modification. Copyright (C) 1999 Elsevier Science B.V.

Original languageEnglish (US)
Pages (from-to)155-160
Number of pages6
JournalGenetic Analysis - Biomolecular Engineering
Volume15
Issue number3-5
DOIs
StatePublished - Nov 1 1999

Fingerprint

Carboxylation
Coagulants
Factor IX
Bioreactors
Human Mammary Glands
Human engineering
Protein C
Recombinant Proteins
Glutamic Acid
Swine
1-Carboxyglutamic Acid
Proteins
Acids
Post Translational Protein Processing
Agriculture
Conformations
Amino Acids
Peptides
Polypeptides
Protein Sequence Analysis

Keywords

  • Mammary gland
  • Protein C
  • Transgenic pigs

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Genetics

Cite this

Transgenic pigs as bioreactors : A comparison of gamma-carboxylation of glutamic acid in recombinant human protein C and factor IX by the mammary gland. / Van Cott, Kevin; Butler, Stephen P.; Russell, Christopher G.; Subramanian, Anuradha; Lubon, Henryk; Gwazdauskas, F. C.; Knight, James; Drohan, William N.; Velander, William H.

In: Genetic Analysis - Biomolecular Engineering, Vol. 15, No. 3-5, 01.11.1999, p. 155-160.

Research output: Contribution to journalArticle

Van Cott, Kevin ; Butler, Stephen P. ; Russell, Christopher G. ; Subramanian, Anuradha ; Lubon, Henryk ; Gwazdauskas, F. C. ; Knight, James ; Drohan, William N. ; Velander, William H. / Transgenic pigs as bioreactors : A comparison of gamma-carboxylation of glutamic acid in recombinant human protein C and factor IX by the mammary gland. In: Genetic Analysis - Biomolecular Engineering. 1999 ; Vol. 15, No. 3-5. pp. 155-160.
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