The Effects of Storage and Heparin on the Activity of Serum Complement, with Particular Reference to the Detection of Blood Group Antibodies

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Abstract
Normal sera were stored at -90 C., -55 C., -20 C.room temperature (20 to 24 C.), and 37 C. from 24 hr. to 3 months. Complement activities of the stored sera were measured by hemolysis of sensitized sheep erythrocytes and by an antiglobulin test assay. The antiglobulin test assay tested the ability of the stored sera to act as a source of complement in the two-stage EDTA antiglobulin test of Polley and Mollison,18 using human erythrocytes sensitized with nonagglutinating IgM Lewis antibody. Hemolytic assays closely paralleled antiglobulin assays. At levels below 60% complement activity there was a danger of missing weak complement-binding antibodies. An average of these assays showed that at 37 C. activity fell to 30% in 24 hr.; at room temperature activity was 40% at 48 hr., zero at 72 hr.; at 4 C. it was 90% at 72 hr. and 60% at 2 weeks. At -20 C. activity was more than 60% for 2 months, and at -55 C. and -90 C. more than 90% activity was retained at 3 months. Heparin was found to be anticomplementary when more than 5 units (0.05 mg.) of heparin per ml. of serum were present. More than 50 units (5 mg.) of heparin per ml. of serum were necessary before weak complement-binding antibody was missed completely by the antiglobulin test.

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