Development of an Immunoassay for Bacterial Endotoxins
Annual rept. 15 Apr 1982-14 Apr 1983
SCRIPPS CLINIC AND RESEARCH FOUNDATION LA JOLLA CA
Pagination or Media Count:
This report describes 1 progress in establishing and validating an immunoassay for lipopolysaccharides and 2 steps taken to begin to characterize the antigenic site of LPS which is involved in the assay. In step 1 of the assay polystyrene tubes are coated with Re595-LPS, washed with bovine serum albumin solution, and dried. In step 2 the sample to be assayed is mixed with I125-labelled immunopurified anti-Re595-LPS IgG and allowed to form soluble antigen-antibody complexes. The mixture is transferred to an LPS coated tube, the uncomplexed I125-IgG binds to the tube, and the soluble LPS-antibody complexes are washed away. Finally in step 3 the LPS coated tubes with I125- IgG bound to them are conducted to determine the I125 content and thus the amount of bound IgG. These data are then used to prepare a standard curve or to calculate the concentration of LPS in a sample.