Monoclonal antibodies to human C4. I. Purification of hemolytically active C4 from small volumes of normal serum.
A C4-specific monoclonal antibody has been developed that allows the rapid purification of C4 from whole serum or plasma by affinity chromatography. 1- to 2-ml volumes of serum are loaded onto a small column of antibody-coupled agarose beads and washed through with buffer. C4 is eluted by adjusting the pH to 11.2. The purified C4 is free of extraneous proteins as detected by SDS-polyacrylamide gel electrophoresis and retains high activity as assessed by hemolytic assay and incorporation of [14C]-methylamine. Columns are reusable, and the entire procedure can be adapted for large-scale purifications.
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002846:Chromatography, Affinity / Q000379:methods; D003181:Complement C4 / Q000276:immunology* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D006461:Hemolysis; D006801:Humans; D007163:Immunosorbent Techniques
D000911:Antibodies, Monoclonal; D003181:Complement C4