Large-scale production and properties of immunoaffinity-purified human activated protein C concentrate.
Activated protein C (APC) is a highly specific serine proteinase which functions as an important naturally occurring antithrombotic enzyme. APC also has anti-inflammatory properties. We have developed a large-scale process for the production of APC for therapeutic use starting with cryoprecipitate-poor human plasma. This report describes the process, its performance at the pilot plant scale, and the characteristics of immunoaffinity-purified human APC concentrate referred to as APC (human). The process consists of three chromatographic steps, an enzymatic conversion step, and incorporates a solvent/detergent treatment step for the inactivation of lipid-enveloped viruses. Solvent/detergent was shown to rapidly inactivate spiked HIV-1, as well as three marker viruses to nondetectable levels under process conditions. The immunoaffinity-purified protein C (PC) intermediate was enriched 13,600-fold over plasma and had a specific activity of 231 U/mg. The overall yield of the process following enzymatic conversion of the PC intermediate to APC and its processing by anion exchange chromatography was 36%. APC (human) was shown to be highly purified, functional and stable.
Orthner
C L
CL
Jerome H. Holland Laboratory, American Red Cross, Rockville, Md, USA.
Ralston
A H
AH
Gee
D
D
Kent
R
R
Kolen
B
B
McGriff
J D
JD
Drohan
W N
WN
D000911:Antibodies, Monoclonal; D001779:Blood Coagulation Factors / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D006801:Humans; D007700:Kinetics; D011486:Protein C / Q000302:isolation & purification*
D000911:Antibodies, Monoclonal; D001779:Blood Coagulation Factors; D011486:Protein C