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Preparation of monoclonal antibody to streptococcal nephritis strain associated protein

Journal of Nippon Medical School 52(5): 530-536

Preparation of monoclonal antibody to streptococcal nephritis strain associated protein

The extracellular protein (ECP) of group A streptococcal strains isolated from patients with acute post-streptococcal glomerulonephritis possesses a protein (mol. wt. 46K daltons) which was assumed to be specific to these strains. This protein was designated as the nephritis-strain associated protein (NSAP). In order to detect the NSAP in ECP, monoclonal antibody (MoAb) to NSAP is useful. BALB/c mice were immunized three times with ECP containing the NSAP intraperitoneally. Spleen cells were prepared from immunized mice three days after the final immunization and fused to myeloma cells (P-3-NS-1-Ag 4-1) using polyethylene glycol. The antibody of the NSAP in culture supernatants of hybridoma cells was analyzed by ELISA. Four clones producing the antibody to the NSAP were obtained and the antibodies belonged to the IgM or IgG classes. It was confirmed by using the nitrocellulose blotting test of combination with the isoelectric focusing and polyacrylamide gel gradient electrophoresis that the MoAb reacted with the NSAP specifically. The existence of the NSAP in ECP was proved by the sandwich method of ELISA using the peroxidase-labeled MoAb.

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Accession: 006173522

PMID: 3905843

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