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Monoclonal antibodies to sea cucumber polysaccharide and their use in a sandwich ELISA assay.

Antigen was synthesized with L-SCP, a sea cucumber polysaccharide isolated from Pearsonothuria graeffei (Semper) and bovine serum albumin (BSA) as a carrier protein. Spleen cells with high titer antibody producing ability were removed and fused with myeloma cells of SP2/0-Ag14 origin. Three stable murine monoclonal antibodies (MAb ascites) producing cell lines to L-SCP were generated according to a conventional immunization protocol. Their epitope mapping and binding specificity, which was characterized by blocking and inhibition enzyme-linked immunosorbent assay (ELISA) indicated that these specific MAb ascites have similar binding patterns. A sandwich ELISA was developed on the basis of employing L-SCP specific antibodies including MAb 3G6 as capture antibody and HRP-3G6 as detection antibody. The working range for L-SCP in aqueous solution from this method was 100-10,000 ng/mL with good sensitivity, specificity, and precision (relative standard deviation ≤7.9%). Thus the developed ELISA can be used as a convenient tool for the rapid detection of L-SCP in biological examples in the future.