苦马豆素单克隆抗体的制备与鉴定

doi:10.16431/j.cnki.1671-7236.2017.07.035

Abstract

Abstract:

In this test,BALB/c mice were immunized by SW-OVA for the preparation of monoclonal antibody against swainsonine (SW). Hybridoma cells that could secrete specific antibody against SW were prepared by hybridoma technique,and then the strain that secreted monoclonal antibody against SW designated 1F10 was prepared,and the chromosome average number of 1F10 cell was 45 to 50 couples. The ELISA titers of cell supernatant were 1:25 600, and that of ascites were 1:80 000.The subclasses of monoclonal antibody was IgG1,and the affinity constant was 1.14×10¹⁰,the purity of ascites antibodies was up to 98%,and the recovery rate was 80%.The result of sodium dodecyl sulfate polyacrylamide gel electropheresis proved that purified antibody had been obtained that the molecular weight of H-chain and L-chain of antibody was about 50 and 25 ku. The monoclonal antibody against SW specifically bound to SW determined by Western blotting. The linear range was 4 to 128 μg/mL (R²=0.9969) and no cross-reactivity was detected with BSA,gelatin,polylysine,me-Gal,etc. The result laid the foundation for immunodetection on SW and immunological prevention of animal toxic disease.

Key words: swainsonine; monoclonal antibody; hybridoma cell; indirect ElISA

CLC Number:

R392

JIA Qi-zhen, CHEN Gen-yuan, WANG Lian-qun, YASEN·Maimaiti, WANG Shuai. Preparation and Identification of Monoclonal Antibody Against Swainsonine[J]. , 2017, 44(7): 2155-2159.

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Preparation and Identification of Monoclonal Antibody Against Swainsonine

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