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Activation of recombinant human protein C
T K Lee1, N Bangalore, W Velander
1Holland Laboratory, American Red Cross, Rockville, MD 20855, USA.
Thrombosis Research
|May 1, 1996
Summary
Researchers created genetically modified pigs to produce recombinant human Protein C (rHPC). The study found rHPC showed similar biological activity to natural human Protein C when activated by specific enzymes.
Area of Science:
- Biotechnology
- Biochemistry
- Genetics
Background:
- Protein C is a crucial anticoagulant protein.
- Recombinant protein production in transgenic animals offers a potential source for therapeutic proteins.
- Understanding the activation and activity of recombinant proteins is vital for their application.
Purpose of the Study:
- To produce recombinant human Protein C (rHPC) in the milk of transgenic swine.
- To compare the activation and biological activity of rHPC with naturally occurring human Protein C (HPC).
Main Methods:
- Production of rHPC in transgenic swine milk.
- Purification of rHPC.
- Analysis of rHPC interaction with Protac, thrombin/thrombomodulin, and thrombin alone.
- Assessment of amidolytic and anticoagulant activities.
Main Results:
- Recombinant activated Protein C (rAPC) exhibited approximately 80% of the amidolytic and anticoagulant activities of its plasma-derived counterpart after Protac activation.
- Both natural and recombinant activated Protein C showed similar enzymatic and biological activities upon activation by the thrombin/thrombomodulin complex.
- Differences in activator mechanisms and structural variations between HPC and rHPC may explain activity discrepancies.
Conclusions:
- Transgenic swine can be utilized for the production of functional recombinant human Protein C.
- rHPC demonstrates comparable biological activity to native HPC, particularly when activated by the thrombin/thrombomodulin complex.
- Further investigation into activator mechanisms and protein structure is warranted.