A simple method for isolating specific antibodies to complement components.
A simple procedure for isolating specific antibodies to complement components C3, C4 and C5b-9 from whole rabbit antisera or immunoglobin preparations is described. Antisera are first reacted with complement-treated sheep erythrocyte membranes. Immunoglobulins are eluted from the washed membranes with acetic acid at pH 2.4, and further purified by one absorption/desorption step on Protein A Sepharose. The final protein preparations contain greater than 90% rabbit immunoglobins, as assessed by quantitative immunoelectrophoresis and SDS polyacrylamide gel electrophoresis. Binding assays with radioiodinated protein preparations indicate a content of specific antibodies in the range 5-55% of total protein. Since there is virtually no non-specific binding of protein to control cells or membranes, the immunoglobulin preparations are well suited for use in sensitive immunoassays for complement components.
D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D000918:Antibody Specificity; D003165:Complement System Proteins / Q000276:immunology*; D004910:Erythrocyte Membrane / Q000276:immunology; D006801:Humans; D007074:Immunoglobulin G / Q000302:isolation & purification; D007158:Immunologic Techniques; D011817:Rabbits; D012756:Sheep
D000906:Antibodies; D007074:Immunoglobulin G; D003165:Complement System Proteins