The development and interlaboratory validation of a quantitative antibody capture ELISA for the measurement of specific guinea-pig IgG1: an alternative to the passive cutaneous anaphylaxis assay

By Babcock, L.S. - Kawabata, T.T. - Moore, V.S. - Condo, C.M. - Prento, A.

Specific guinea-pig IgG1 has traditionally been measured by using an in vivo guinea-pig passive cutaneous anaphylaxis (PCA) assay. This paper describes the development and validation of a quantitative enzyme-linked immunosorbent assay (ELISA) for specific IgG1 against two protein antigens (Alcalase and Enzyme B) as an alternative to the PCA assay. The ELISA format involved a rabbit antibody bound to microtitre plates to capture the antigen. The test sera is added to this, followed sequentially by goat anti-guinea-pig IgG1 and rabbit antigoat-IgG-alkaline phosphatase conjugate. Aliquots of each serum sample from immunised guinea-pigs were analysed with the ELISA for specific IgG1 titres in three laboratories and were compared to titres determined by using the PCA assay. The findings demonstrate that there is a good correlation between the ELISA and the PCA assay and that the ELISA shows good interlaboratory reproducibility. Thus, the antibody capture ELISA described in this report is a valid and robust replacement for the guinea-pig PCA assay.