Fractionation of human antibody to factor VIII:C: and IRMA for phospholipid binding sites on factor VIII C:Ag

Br J Haematol. 1984 Aug;57(4):633-6. doi: 10.1111/j.1365-2141.1984.tb02940.x.

Abstract

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.

MeSH terms

  • Antibodies / immunology*
  • Antigens / analysis
  • Antigens / immunology*
  • Binding Sites, Antibody
  • Factor VIII / analysis
  • Factor VIII / immunology*
  • Humans
  • Immunoglobulin Fab Fragments
  • Immunosorbent Techniques
  • Phospholipids / immunology
  • Radioimmunoassay

Substances

  • Antibodies
  • Antigens
  • Immunoglobulin Fab Fragments
  • Phospholipids
  • Factor VIII