Evaluation of complement activity by an enzyme immunoassay.
An ELISA-type assay useful for the evaluation of the complement activity in serum is described. Aggregated pooled human IgG (IgGn) prepared so as to exclude large and small aggregates, to resemble soluble circulating immune complexes, was used to coat polystyrene microwells to serve as initiator of complement activation. Fresh serum, at different dilutions, as the source of the complement to be evaluated was added and the plate incubated 90 min at 37 degrees C. Then, a peroxidase-labeled antihuman C3c antibody was added to react with the bound C fragments. This was followed by 2,2'-azino-di-3-ethyl benzothiazoline sulfonic acid (ABTS), as the color reagent used for detection of the enzyme activity. In this system, theoretically, the levels of activating and regulatory complement components are evaluated up to the level of C3 splitting. The assay was applied in healthy volunteers to set normal values and in 15 patients suffering from systemic lupus erythematosus making possible the differentiation of those with normal and low complement levels.
Rossi Devivo
M L
ML
Laboratory of Pathophysiology, Instituto Venezolano de Investigaciones Científicas (IVIC), Caracas.
Romano
E L
EL
Suárez
G
G
Rodríguez
M
M
Blasini
A M
AM
Soyano
A
A
D003167:Complement Activation; D003176:Complement C3 / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005260:Female; D006461:Hemolysis; D006801:Humans; D005779:Immunodiffusion / Q000379:methods; D007074:Immunoglobulin G / Q000378:metabolism; D008180:Lupus Erythematosus, Systemic / Q000097:blood / Q000276:immunology; D008297:Male; D011489:Protein Denaturation; D015203:Reproducibility of Results; D013997:Time Factors
D003176:Complement C3; D007074:Immunoglobulin G; D003165:Complement System Proteins