Enzyme Immunoassay for N^α-(N-Acetylmuramylalanyl-D-isoglutaminyl)-N^ε-stearyllysine

概要

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An enzyme immunoassay method for N^α-(N-acetylmuramylalanyl-D-isoglutaminyl)-N^ε-stearyllysine (MDP-Lys-L18) was established. A heterologous system was adopted based on the enzyme-labeled conjugate (EC) prepared by linking MDP-lysine (MDPL ; an analogue of MDP-Lys-L18) to horseradish peroxidase (HRP) (EC1.11.1.7) by the sodium metaperiodate method, and anti-MDP-Lys-L18 antiserum obtained from a rabbit that had been immunized with MDP-Lys-L18 conjugated with bovine serum albumin (BSA) by the carbodiimide method. The bound and free ECs were separated by a double antibody method. The lower limit for the quantitative determination of MDP-Lys-L18 in serum was estimated to be 100 pg/ml and the coefficients of variation of this assay (intra-assay) were between 3 and 8%. This enzyme immunoassay is superior in sensitivity to the radioimmunoassay previously reported. Serum concentrations of MDP-Lys-L18,after subcutaneous administration at a dose of 50μg/kg to rats, were determined by means of this assay.
1985-12-25