[A method of isolating C1q from human serum and its use in the solid-phase immunoenzyme determination of immune complexes].
C1q was isolated from human serum by dialysis in 0.24 M EDTA, followed by affinity chromatography on immobilized IgG and removal of IgG traces in a column with anti-IgG antibodies. Microplates were coated with C1q in PBS at 10-20 mg/l, nonspecific binding sites were saturated with human serum albumin. The sera were diluted 16-fold in 0.05 M PBS, 0.01 M EDTA, 0.05% Tween. After incubation with diluted samples the plates were treated with horseradish peroxidase--anti-human IgG conjugates. Enzymic activity was measured by adding p-phenylenediamine (0.2 g/l) in acetate buffer, pH 5.9, containing 0.05% H2O2. The sensitivity of the assay ranged between 2.5 and 300 mg/l.
Belozerov
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AP
D000936:Antigen-Antibody Complex / Q000032:analysis*; D003166:Complement Activating Enzymes / Q000302:isolation & purification*; D003172:Complement C1 / Q000302:isolation & purification*; D015922:Complement C1q; D006801:Humans
D000936:Antigen-Antibody Complex; D003172:Complement C1; D015922:Complement C1q; D003166:Complement Activating Enzymes
Metod vydeleniia C1q iz syvorotki cheloveka i ispol'zovaniia ego v tverdofaznom immunofermentnom opredelenii immunnykh kompleksov.