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

Specific guinea-pig IgG1 has traditionally been measured by using an i n vivo guinea-pig passive cutaneous anaphylaxis (PCA) assay. This pape r describes the development and validation of a quantitative enzyme-li nked 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 sequ entially by goat anil-guinea-pig IgG1 and rabbit anti-goat-IgG-alkalin e 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 inte rlaboratory reproducibility. Thus, the antibody capture ELISA describe d in this report is a valid and robust replacement for the guinea-pig PCA assay.