A monoclonal antibody to VIII:C produced by a mouse hybridoma
HP Muller, NH van Tilburg, J Derks, E Klein-Breteler and RM Bertina
Spleen cells of a BALB/c mouse immunized with factor VIII procoagulant
activity (VIII:C) (isolated by affinity chromatography) were fused with
mouse myeloma cells (P3 x 63 Ag8). After the fusion 12/32 wells produced an
inhibitor to VIII:C. Cells from one well (1B3) were subcloned four times in
order to isolate the hybridoma that produces the anti-VIII:C antibody.
Injection of hybridoma cells in pristane pretreated BALB/c mice results in
anti-VIII:C titers of 5000-10,000 Bethesda U/ml. Analysis of the produced
immunoglobulin demonstrated heavy chains of IgG1 (produced by the myeloma
cell line) and IgG2b subclass. The 1B3 antibody neutralizes VIII:C in LMW
FVIII, crysosupernatant, cryoprecipitate, and normal plasma. It was found
that binding of the IgG to FVIII results in a delay in its activation and
not in an inhibition of its cofactor activity. The antibody removes VIII:C
from pooled normal plasma when coupled to Sepharose; when coupled to
plastic tubes, it binds VIIICAG from isolated VIII:C, purified FVIII, and
pooled normal plasma; it does not bind VIIIR:AG, fibrogen, or serum
VIIICAG. The 1B3 antibody can be used successfully in an IRMA for VIIICAG.
Volume 58,
Issue 5,
pp. 1000-1006,
11/01/1981
Copyright © 1981 by The American Society of Hematology
