Preparation of an R3 reagent (serum depleted of the third component of human complement (C3)) by immunoadsorption. Application to the hemolytic assay of human C3.
We have devised a simple one-step preparation of C3 depleted serum (R3). Fresh normal human serum, with complement activation inhibitors, was depleted of C3 by affinity chromatography on a Sepharose anti-C3c. To prevent non-specific interactions, a high ionic strength buffer containing saccharose was used. Immunochemical analysis and complement activity assays demonstrate that this C3 depleted serum is a suitable R3 reagent. This R3 reagent has been used successfully for titration of hemolytic activity of C3 in normal and abnormal sera. Two simple, specific and sensitive assays are described.
D000818:Animals; D001770:Blood Bactericidal Activity; D003172:Complement C1; D003176:Complement C3 / Q000172:deficiency*; D003181:Complement C4; D003182:Complement C5; D004926:Escherichia coli; D006461:Hemolysis; D006801:Humans; D007163:Immunosorbent Techniques; D008837:Micrococcus; D008970:Molecular Weight; D009895:Opsonin Proteins; D011817:Rabbits; D012756:Sheep
D003172:Complement C1; D003176:Complement C3; D003181:Complement C4; D003182:Complement C5; D009895:Opsonin Proteins