title: Monoclonal antibodies to sea cucumber polysaccharide and their use in a sandwich ELISA assay.
abstract: 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.
authors.affiliation: School of Life Science and Technology, China Pharmaceutical University, Nanjing, P.R. China .
authors.forename: Junjun
authors.initials: J
authors.lastname: Wang
authors.forename: Hu
authors.initials: H
authors.lastname: Zhao
authors.forename: Ying
authors.initials: Y
authors.lastname: Zheng
authors.forename: Wei
authors.initials: W
authors.lastname: Liu
authors.forename: Hua
authors.initials: H
authors.lastname: Zhou
authors.forename: A-ru-na
authors.initials: AR
authors.lastname: Han
authors.forename: Xiangdong
authors.initials: X
authors.lastname: Gao
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism*; D000915:Antibody Affinity; D000918:Antibody Specificity; D001202:Ascitic Fluid / Q000378:metabolism; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D047332:Holothuria; D006735:Horseradish Peroxidase / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism*; D007075:Immunoglobulin M / Q000737:chemistry / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D011134:Polysaccharides / Q000737:chemistry / Q000276:immunology*; D015203:Reproducibility of Results; D017186:Titrimetry
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D011134:Polysaccharides; D006735:Horseradish Peroxidase