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膝沟藻毒素GTX1-4单克隆抗体的制备与鉴定

Preparation and identification of monoclonal antibody against Gonyautoxins GTX1-4

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【作者】 黄柏强张卓江天久

【Author】 HUANG Bai-qiang;ZHANG Zhuo;JIANG Tian-jiu;Key Laboratory of Eutrophication and Red Tide Prevention of Guangdong Higher Education Institutes, Research Center of Harmful Algae and Marine Biology, Jinan University;

【通讯作者】 江天久;

【机构】 暨南大学赤潮与海洋生物学研究中心水体富营养化与赤潮防治广东省高校重点实验室

【摘要】 研究膝沟藻毒素GTX1-4单克隆抗体的制备方法。利用甲醛法合成GTX1-4人工抗原,通过小鼠免疫和单克隆抗体技术制备GTX1-4单克隆抗体。结果显示,合成了免疫抗原GTX1-4-BSA和检测抗原GTX1-4-KLH,其偶联比分别为10.7∶1与22.4∶1。筛选出3株能稳定分泌GTX1-4单克隆抗体的杂交瘤细胞1-A4、5-E11和5-A11。5-E11、1-A4诱生腹水单克隆抗体的亚型分别为IgG1和IgG2a,效价均达到64000;5-A11诱生腹水单克隆抗体的亚型为IgG1,效价达到128000。5-A11诱生腹水单克隆抗体的灵敏度为6.25μg/mL,特异性较强。本实验成功合成了GTX1-4人工抗原,并获得了能够稳定分泌GTX1-4单克隆抗体的杂交瘤细胞株。

【Abstract】 The goal of this study was to develop a method for preparation of monoclonal antibody against Gonyautoxins GTX1-4. GTX1-4 artificial antigen was synthesized by formaldehyde method, and GTX1-4 monoclonal antibody was prepared by mouse immunity and monoclonal antibody technology. The results showed that the conjugated ratios of immunizing antigen GTX1-4-BSA and detected antigen GTX1-4-KLH were 10.7∶1 and 22.4∶1, respectively. Three hybridomas cell lines, 1-A4, 5-A11 and 5-E11, which could stably secret GTX1-4 monoclonal antibody were screened. The subtypes of monoclonal antibodies in ascites induced by 5-E11 and 1-A4 were IgG1 and IgG2 a, and the titers of them were up to 64000. The subtype of monoclonal antibody in ascites induced by 5-A11 was IgG1, and its titer could be 128000. The sensitivity of monoclonal antibody in ascites induced by 5-A11 was 6.25 μg/mL, and this antibody has a strong specificity. In the study, consequently, GTX1-4 artificial antigen was successfully synthesized, and the hybridomas cell lines which could stably secret GTX1-4 monoclonal antibody was obtained.

【基金】 科技部科技基础资源调查专项项目(2018FY100201);国家重点研发计划项目(2019YFC1407900);国家自然科学基金面上项目(41276091);海洋公益性行业科研专项项目(201305010)
  • 【文献出处】 海洋环境科学 ,Marine Environmental Science , 编辑部邮箱 ,2021年03期
  • 【分类号】X56;Q503
  • 【下载频次】138
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