TY - JOUR
T1 - Transgenic pigs as bioreactors
T2 - A comparison of gamma-carboxylation of glutamic acid in recombinant human protein C and factor IX by the mammary gland
AU - Van Cott, Kevin E.
AU - Butler, Stephen P.
AU - Russell, Christopher G.
AU - Subramanian, Anu
AU - Lubon, Henryk
AU - Gwazdauskas, F. C.
AU - Knight, James
AU - Drohan, William N.
AU - Velander, William H.
N1 - Funding Information:
We are grateful to Dr Barry Williams for assistance with animal husbandry. This work was supported from grants from the American Red Cross.
PY - 1999/11
Y1 - 1999/11
N2 - 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.
AB - 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.
KW - Mammary gland
KW - Protein C
KW - Transgenic pigs
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U2 - 10.1016/S1050-3862(99)00020-0
DO - 10.1016/S1050-3862(99)00020-0
M3 - Article
C2 - 10596756
AN - SCOPUS:0032730944
VL - 15
SP - 155
EP - 160
JO - Gene Analysis Techniques
JF - Gene Analysis Techniques
SN - 1871-6784
IS - 3-5
ER -