Enzyme immunoassays for measuring complement-dependent prevention of immune precipitation (PIP) and solubilization of preformed antigen-antibody complexes (SOL).
We have developed simple and sensitive enzyme-based methods for evaluating the ability of serum complement to prevent immune complex precipitation (PIP) or to solubilize preformed immune complexes (SOL). Alkaline phosphatase, serving both as antigen and label, is added to goat IgG anti-alkaline phosphatase antibodies, with serum present throughout the assay (PIP), or added after immune complex formation (SOL). After incubation at 37 degrees C for 1 h followed by centrifugation, the enzyme activity of the supernatant, reflecting the amount of immune complexes in solution, is measured by colorimetry. Results are expressed with reference to a standard serum pool assigned 100 arbitrary units (AU). Intra- and inter-assay variabilities are within 10%. The normal ranges were 67-133 AU for PIP and 72-129 AU for SOL. These methods have been standardized for clinical use in relation to impaired complement function and immune complex disease, and adapted for measuring complement mediated binding of immune complexes to erythrocytes. They are sensitive, easy to perform and do not require expensive facilities. By measuring the interaction of complement with immune complexes, these methods may highlight aspects of the classical and the alternative pathway that are different from those detected using haemolysis as an endpoint.
Department of Immunology, National University Hospital, Landspítalinn, Reykjavík, Iceland. garason@rsp.is
G J
GJ
Arason
M S
MS
D'Ambrogio
T
T
Víkingsdóttir
A
A
Sigfússon
H
H
Valdimarsson
D000469:Alkaline Phosphatase / Q000276:immunology / Q000592:standards; D000936:Antigen-Antibody Complex / Q000097:blood / Q000378:metabolism*; D002498:Centrifugation; D003175:Complement C2 / Q000172:deficiency; D003165:Complement System Proteins / Q000172:deficiency / Q000502:physiology*; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D007105:Immune Complex Diseases / Q000097:blood; D007124:Immunoenzyme Techniques / Q000379:methods* / Q000592:standards; D011233:Precipitin Tests / Q000379:methods / Q000592:standards; D012016:Reference Values; D015203:Reproducibility of Results; D012995:Solubility; D013997:Time Factors
D000936:Antigen-Antibody Complex; D003175:Complement C2; D003165:Complement System Proteins; D000469:Alkaline Phosphatase