Validation of an ELISA for the quantitation of human complement receptor 1

An ELISA was developed and validated for the quantitation of Complement Receptor 1 (CR1) in human plasma. The ELISA employed a monoclonal anti-CR1 antibody adsorbed onto microtiter plates to capture CR1 in human plasma. The captured CR1 was treated with a detecting antibody which had a different epitopic specificity for CR1. HRP conjugated anti IgG (secondary antibody) was used for quantitation. The standard curve covered a wide range from 10 pg to 800 pg. The inter- and intra-assay variation were found to be low and within the acceptable limits. Specificity and accuracy for the assay was established by ensuring negligible cross reactivity with other proteins and an excellent parallelism between the sample and standard curve. The samples were checked for loss of sCR1 levels through freeze/thaw cycles at different intervals of time stored at -70 degrees C.