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Fractionation of human antibody to factor VIII:C: an IRMA for phospholipid binding sites on factor VIII C: Ag
Author(s) -
KemballCook G.,
Barrowcliffe T. W.
Publication year - 1984
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.1365-2141.1984.tb02940.x
Subject(s) - antibody , immunoradiometric assay , phospholipid , chemistry , microbiology and biotechnology , binding site , vesicle , antigen , incubation , fractionation , radioimmunoassay , biochemistry , biology , chromatography , immunology , membrane
S ummary. Labelled Fab’ fragments, derived from the plasma of a severe haemophiliac with antibody directed against factor VIII clotting antigen (VIII C:Ag), were fractionated by immunoabsorption with, first, a complex of phospholipid (PL) vesicles and factor VIII and, second, with factor VIII alone. Two pools of labelled anti‐VIII C:Ag were obtained and were used in immunoradiometric assays (IRMAs) for VIII C:Ag. With one pool (non‐PL‐site antibody) VIII C:Ag assays were unaffected by pre‐incubation of factor VIII with PL vesicles; however, binding of the second pool of antibody to VIII C:Ag was prevented by PL preincubation, indicating that these antibody molecules bind at or near a phospholipid binding site on VIII C:Ag (PL‐site antibody). Assays of VIII C:Ag in an intermediate purity factor VIII concentrate with these two antibody pools indicate that more than one third of the VIII C:Ag may be bound to PL.