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Isolation of Human Antibodies to Factor VIII
Author(s) -
Lavergne J. M.,
Meyer Dominique,
Koutts J.,
Larrieu MarieJosé
Publication year - 1978
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1111/j.1365-2141.1978.tb05839.x
Subject(s) - antibody , sephadex , antigen , chemistry , titer , size exclusion chromatography , microbiology and biotechnology , biochemistry , immunology , medicine , biology , enzyme
S ummary . It has been claimed that human anti‐VIII:C antibodies do not form stable complexes with factor VIII and this fact has hampered in the past the isolation of such antibodies. In this study the purification of human anti‐VIII:C antibodies appearing in haemophiliac patients following replacement therapy has been achieved using two different systems. In a liquid phase system, purified human factor VIII was mixed with IgG from a haemophilic patient with a high titre antibody. Specific anti‐VIII:C antibodies were recovered following filtration of the antigen‐antibody complexes on Biogel A‐5m, dissociation of complexes at pH 3.5 and final isolation by filtration on Sephadex G‐200. In a solid phase system, the same IgG fraction was specifically bound to insolubilized human factor VIII. Purified anti‐VIII:C antibodies were subsequently recovered by elution of antigen‐antibody complexes with magnesium chloride. The results demonstrated that stable complexes form between anti‐VIII:C antibodies and either the whole factor VIII molecule, or VIII:C dissociated by previous interaction with the antibodies. It is postulated that, in vivo , similar antigen‐antibody complexes may form following replacement therapy in haemophilic patients with antibody.