A Soluble Recombinant Factor VIII Fragment Containing the A2 Domain Binds to Some Human Anti-Factor VIII Antibodies that Are not Detected by Immunoblotting

Dorothea Scandella; Lisa Timmons; Mora Mattingly; Norma Trabold; Leon W Hoyer

doi:10.1055/s-0038-1648520

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 1992; 67(06): 665-671
DOI: 10.1055/s-0038-1648520

Original Articles

Schattauer GmbH Stuttgart

A Soluble Recombinant Factor VIII Fragment Containing the A2 Domain Binds to Some Human Anti-Factor VIII Antibodies that Are not Detected by Immunoblotting

Dorothea Scandella

The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross Blood Services, Rockville, MD, USA

,

Lisa Timmons

The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross Blood Services, Rockville, MD, USA

,

Mora Mattingly

The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross Blood Services, Rockville, MD, USA

,

Norma Trabold

The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross Blood Services, Rockville, MD, USA

,

Leon W Hoyer

The Jerome H. Holland Laboratory for the Biomedical Sciences, American Red Cross Blood Services, Rockville, MD, USA

› Author Affiliations

Abstract

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Summary

Human factor VIII (f VIII) inhibitors are pathologic antibodies that inactivate fVIII. A cDNA clone was modified to encode f VIII amino acid residues 373-740 for expression in a baculovirus vector in insect cells. The encoded protein fragment H2 was produced as a soluble, secreted protein, and it was used to test inhibitor plasmas for the presence of antibodies that were not detected by immunoblotting. Seven of 13 inhibitors that bound, only to the fVIII light chain by immunoblotting also bound to fragment H2 in an immunoprecipitation assay. Thus multi-chain inhibitor reactivity of inhibitors is more frequent than previously reported. One of these inhibitors was shown to share the epitope for other inhibitors that bind to H2 within amino acid residues 373-541 in immunoblotting assays. The sensitive immunoprecipitation assay described allows determination of relative H2 binding capacity of the total IgG and epitope localization of inhibitors that cannot be similarly characterized by immunoblotting.

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References

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