DEVELOPMENT OF MONOCLONAL ANTIBODIES AND QUANTITATIVE SANDWICH ENZYME LINKED IMMUNOSORBENT ASSAY FOR THE CHARACTERISTIC SIALOGLYCOPROTEIN OF EDIBLE BIRD'S NEST

作者：Zhang, Shiwei; Lai, Xintian; Liu, Xiaoqing; Li, Yun; Li, Bifang; Huang, Xiuli; Zhang, Qinlei; Chen, Wei; Lin, Lin; Yang, Guowu^*

来源：Journal of Immunoassay and Immunochemistry, 2013, 34(1): 49-60.

DOI：10.1080/15321819.2012.680527

• authentication
• edible bird's nest
• ELISA
• sialoglycoprotein

The article presents a sandwich enzyme linked immunosorbent assay (ELISA) for identification of edible bird's nest. The characteristic sialoglycoproteins were found by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and purified by liquid-phase isoelectric focusing (LIEF). According to the analysis, the molecular weight was 106128kDa and the isoelectric point was pH 3.0. Two anti-characteristic sialoglycoprotein monoclonal antibodies were produced. The monoclonal antibodies were examined by western-blot assay. One of the monoclonal antibody was used as coating and the other as the enzyme-labeled antibody after being coupled to horseradish peroxidase (HRP). Based on the optimized ELISA condition, the method was established with IC50 of 1.5ng/mL, and low cross-reactivity with various fake materials (<0.01%). ELISA provided a suitable means for screening of a large number of samples. The coefficients of variation were between 2.9% and 5.8%.

• 出版日期2013-1-1
• 单位深圳市计量质量检测研究院