Preparation of factor IX deficient human plasma by immunoaffinity chromatography using a monoclonal antibody.
A murine hybridoma clone is described that grows continuously in culture and produces a monoclonal antibody we have called Royal Free Monoclonal Antibody to factor IX No. 1 (RFF-IX/1). This has high affinity for a coagulation site on factor IX. RFF-IX/1 immobilised on sepharose can be used to deplete factor IX from normal human plasma. This immunoaffinity depleted plasma is indistinguishable from severe Christmas disease plasma and can be used as the substrate in a one stage coagulation assay for factor IX. The affinity column has high capacity and can be regenerated so that large scale production from normal plasma of factor IX deficient plasma as a diagnostic reagent is now feasible.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D002846:Chromatography, Affinity; D005164:Factor IX / Q000037:antagonists & inhibitors / Q000276:immunology / Q000302:isolation & purification; D002836:Hemophilia B / Q000097:blood* / Q000175:diagnosis; D006801:Humans; D007202:Indicators and Reagents; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D005164:Factor IX