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Thromb Haemost 1986; 56(03): 271-276
DOI: 10.1055/s-0038-1661665
Original Article
Schattauer GmbH Stuttgart

Characterization of Four Monoclonal Antibodies to Factor VIII Coagulant Protein and Their Use in Immunopurification of Factor VIII

Marie-Pascale Croissant
The Division of Research and Development, Centre National de Transfusion Sanguine, Les Ulis, France
,
Hendrik vande Pol
The Division of Research and Development, Centre National de Transfusion Sanguine, Les Ulis, France
,
Helen H Lee
The Division of Research and Development, Centre National de Transfusion Sanguine, Les Ulis, France
,
Jean-Pierre Allain
The Division of Research and Development, Centre National de Transfusion Sanguine, Les Ulis, France
› Author Affiliations
Further Information

Publication History

Received 29 November 1985

Accepted after revision 22 August 1986

Publication Date:
18 July 2018 (online)

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Summary

Four monoclonal anti-VIII: C antibodies were obtained from the fusion of the splenocytes of one B alb/C mouse with a specific activity ranging from 2.3 to 45,000 U/mg when purified from ascitic fluid. Only one antibody was able to inhibit completely Factor VIII: C in normal plasma. The four antibodies could bind Factor VIII: CAg in plasma and commercial concentrate both in liquid and solid phase, and were suitable for immunopurification of Factor VIII :C.

Three antibodies competed with polyclonal anti-VIII: CAg Fab′ in a liquid phase IRMA, and all of them were able to displace their own binding to Factor VIII: CAg. Competition studies between monoclonal antibodies for the binding to Factor VIII: CAg were performed and showed the recognition of different epitopes and various functional impact. These studies indicate that at least one antibody, with the lowest anti-VIII:C titer clearly recognizes a different epitope of VIII: C than those recognized by the others. Affinity constants ranged from 109 to 1010 l/mole.

Key words

Monoclonal antibodies - Factor VIII: C - Antibody affinity - Competition studies - Immunopurification
 

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