抗酸土脂环酸芽孢杆菌(Alicyclobacillus acidoterrestris)单克隆抗体制备

【背景】基于本实验室已经建立的脂环酸芽孢杆菌检测和鉴定方法工作基础之上,以期建立具有很好的经济价值和实用价值,且更为方便、快捷、准确、特异、灵敏的检测方法。【目的】实现对果汁生产中脂环酸芽孢杆菌从原料到成品的快速检测和鉴定。【方法】采用杂交瘤细胞技术,以Alicyclobacillus acidoterrestris(ATCC49025)免疫BALB/c小鼠,用建立的间接ELISA方法筛选杂交瘤细胞,得到了3株能稳定分泌A.acidocaldarius抗体的杂交瘤细胞株,其中两株为Ig G1亚类,并对其进行生物学特性的鉴定。【结果】得到的两株单抗3F7和9C4是针对不同的抗原位点,且多次传代后稳定性基本保持不变;特异性实验表明两株单抗均不与A.acidocaldarius(NCIMB11725)、Bacillus cereus(ATCC11778)、Bacillus subtilis(ATCC11774)、A.cycloheptanicus(ATCC49029)等发生交叉反应。【结论】3F7和9C4这两株单抗可以进一步用于检测胶体金试纸条的研制。

Preparation of monoclonal antibodies against Alicyclobacillus acidoterrestris

[Background] This project is based on the methods of detection and identification of Alicyclobacillus in our laboratory. We hope to establish a good economic value and practical value, more convenient, fast, accurate, specific and sensitive detection methods. [Objective] In order to detect and identify Alicyclibacillus rapidly in raw materials or finished products in fruit juice production enterprises. [Methods] We immunized BALB/c mice with A. acidoterrestris (ATCC49025) and established indirect ELISA to screen hybridoma cells. using hybridoma technology, we obtained 3 hybridoma cell lines that could stably secrete A. acidoterrestris antibody. Two of the three were IgG1 and their biological characteristics were identified. [Results] Two monoclonal antibodies 3F7 and 9C4 had different antigenic sites and the stability remained unchanged after multiple passages. Specificity experiments showed that the two monoclonal antibodies did not cross react with A. acidocaldarius (NCIMB11725), Bacillus cereus (ATCC11778), Bacillus subtilis (ATCC11774) and A. cycloheptanicus (ATCC49029). [Conclusion] These two monoclonal antibodies can be further used in the development of colloidal gold test strips.