Establishment of monoclonal antibodies against excretory/secretory antigen of Clonorchis sinensis and development of a quantitative double-antibody sandwich ELISA for ESA
ZHU Mu-chun1, LIU Jin-feng2, LU Zhong-kui2, KONG De-long1
1. Department of Pathogen Biology and Immunology, Xuzhou Medical University, Xuzhou 221004, China; 2. Jiangsu Key Laboratory of Immunity and Metabolism, Nanjing 211166, China
Abstract:We aimed to screen the monoclonal antibody(McAb) against excretory/secretory antigen (ESA) of Clonorchis sinensis(C. sinensis) and develop a double anti-body sandwich ELISA for ESA. BALB/c mice were immunized with ESA of C. sinensis, the spleen cells of immunized mice were fused with myeloma cell line SP2/0. Then the hybridoma cell lines secreting high titer of McAbs were screened. The ESA specific McAbs were paired to establish a double antibody sandwich ELISA method. The performance of the double anti-body sandwich ELISA was evaluated by specificity and sensitivity. Two monoclonal antibodies B7 and F1 were selected to establish double antibody sandwich ELISA for ESA of C. sinensis. The immunological subtypes of McAb were identified as IgG1. The detection 1inearity fell in the range of 0.014-0.248 μg/mL, (R2=0.996 9). The detection limit of the assay was 0.014 μg/mL.
朱慕春, 刘劲峰, 陆忠奎, 孔德龙. 华支睾吸虫排泄分泌抗原McAb的制备及双抗夹心ELISA的初步建立[J]. 中国人兽共患病学报, 2018, 34(10): 873-877.
ZHU Mu-chun, LIU Jin-feng, LU Zhong-kui, KONG De-long. Establishment of monoclonal antibodies against excretory/secretory antigen of Clonorchis sinensis and development of a quantitative double-antibody sandwich ELISA for ESA. Chinese Journal of Zoonoses, 2018, 34(10): 873-877.
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