Citations to this article

Heterogeneity of human factor V deficiency. Evidence for the existence of antigen-positive variants
Hui Chong Chiu, … , Eugene Whitaker, Robert W. Colman
Hui Chong Chiu, … , Eugene Whitaker, Robert W. Colman
Published August 1, 1983
Citation Information: J Clin Invest. 1983;72(2):493-503. https://doi.org/10.1172/JCI110997.
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Research Article

Heterogeneity of human factor V deficiency. Evidence for the existence of antigen-positive variants

Abstract

Functional human Factor V has been purified using a rapid immunoaffinity method. Following barium citrate adsorption of plasma, Factor V was precipitated with polyethylene glycol at a concentration between 5 and 14%. The resulting preparation was applied to a column containing an immobilized immunoadsorbent consisting of an IgG fraction containing a naturally occurring human monoclonal (IgG4λ) antibody with inhibitory activity against human Factor V. The solid phase immunoglobulin quantitatively bound Factor V from human plasma. The bound Factor V was effectively eluted with a Tris buffer pH 7.2 containing 1.2 M NaCl and 1 M α-methyl-D-mannoside. The isolated native Factor V with high specific activity (92 U/mg) showed a single band (Mr, 350,000) on both reduced and nonreduced sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Factor V was purified 5,100-fold over plasma with an overall yield of 77%. The purified Factor V when subjected to thrombin activation exhibited an 18-fold increase in coagulant activity.

Authors

Hui Chong Chiu, Eugene Whitaker, Robert W. Colman

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Citations to this article (30)

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