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Abstract
Seven monoclonal antibodies (MAbs) specific for marine fungal polysaccharide YCP isolated from the Phoma herbarum YS4108 were obtained after immunization of BALB/c mice with the conjugate of YCP coupled to bovine serum albumin (BSA). Their epitope mapping and binding specificity characterized by blocking and inhibition Enzyme-linked immunosorbent assay (ELISA) indicated that these specific MAbs have similar binding patterns. An immuno-capture ELISA has been developed on the basis of employing the MAbs N3F and P12B specific for YCP as the capture antibody and the detecting antibody, respectively. The working range for YCP in aqueous solution was 1–10,000 ng/ml with a good reproducibility (relative standard deviation ≤7.2%). Subsequently, the extraction process of YCP from Phoma herbarum YS4108 was successfully optimized by the method.
| ABBREVIATIONS | ||
| MAb: | = | monoclonal antibody |
| ELISA: | = | enzyme-linked assay |
| RSD: | = | relative standard deviation |
| BSA: | = | bovine serum albumin |
| PBS: | = | phosphate-buffered salin |
| FCS: | = | fetal calf serum |
| OPD: | = | o-phenylenediamine |
| HRP: | = | horseradish peroxidase |
| IC50: | = | 50% inhibition |