糖化酶单抗的研制及其抗原识别位点的分析

Abstract

Abstract: To develop a detection method for glucoamylase in honey, BALB/c mice were immunized with the glucoamylase from Aspergillus niger. Splenocytes of the immunized mice were fused with SP2/0 cells with PEG. 12 monoclonal antibodies(McAbs) against the glucoamylase were obtained and identified. 10 hybridoma cell lines represented subtypes IgG1 and the other were IgG2b, and their light chains were κ chain. Western blotting analysis showed that the 12 McAbs specifically recognized glucoamylase. Hybridoma 2H4F9,6H9D8,8F2F11,8F2E9,1A8G6,1C4D5 excreting anti-glucoamylase McAbs were used to be injected into abdominal cavity of mice(ICR). The titers of the McAbs were all above 1∶1×10⁴. Six strains of monoclonal antibodies were confirmed to be specific for four different kinds of epitopes on glucoamylase competitive binding assay. Among them, McAb-6H9D8 and McAb-8F2F11 might direct to the same antigenic determinant on the GA, McAb-1A8G6 and McAb-1C4D5 to the second antigenic site, McAb-8F2E9 to the third antigenic site, McAb-2H4F9 to the fourth antigenic site.

Key words: glucoamylase; monoclonal antibodies; epitopes

CLC Number:

S852.4^＋3

ZHANG Dan, JIANG Wei, LI Shuang-xi, WANG Hui-ze, FENG Yun-fei, LIU Yun-ying, LI Dong-ming, HE Xiang-lai, WANG Han-dong. The Development and Antigenic Analysis of Monoclonal Antibodies to Glucoamylase[J]. , 2012, 39(11): 39-42.

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The Development and Antigenic Analysis of Monoclonal Antibodies to Glucoamylase

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