Preparation and characterization of monoclonal antibodies against a polysaccharide from Flammulina velutipes mycelium.
Antigen was synthesized for the preparation of monoclonal antibodies (MAbs) against FVP2, a polysaccharide from Flammulina velutipes. FVP2 was coupled to bovine serum albumin (BSA) by 1-cyano-4-dimethyl-aminopyridinium tetrafluoroborate (CDAP) to produce a neoglycoprotein with the degree of substitution (DS; approximately 2.5:1 mol of FVP2 hapten/mol of BSA carrier), whose immunogenicity was enhanced in mice. The neoglycoprotein induced vigorous primary IgM and booster IgG responses to FVP2. IgG1 was elicited as the dominant isotype antibody in BALB/c mice. Then, the MAbs to FVP2 were prepared using hybridoma technology. Resulting hybridomas producing anti-FVP2 antibodies were screened by indirect enzyme-linked immunosorbent assay (ELISA). As a result, three hybridoma cell lines named B11D, C8G, and I6D were acquired, and the immunological characteristics of the MAbs were determined.