食品科学 ›› 2019, Vol. 40 ›› Issue (2): 110-116.doi: 10.7506/spkx1002-6630-20180103-029

• 生物工程 • 上一篇    下一篇

抗牛免疫球蛋白G单克隆抗体的制备及鉴定

王丽威1,2,刘金1,武俊瑞1,乌日娜1,岳喜庆1,*   

  1. (1.沈阳农业大学食品学院,辽宁?沈阳 110866;2.辽宁工程技术大学理学院,辽宁?阜新 123000)
  • 出版日期:2019-01-25 发布日期:2019-01-22
  • 基金资助:
    2011年度国家星火计划项目(2011GA650001);“十二五”农村领域国家科技计划课题(2013BAD18B03-02)

Preparation and Identification of Monoclonal Antibody against Bovine Immonoglobulin G

WANG Liwei1,2, LIU Jin1, WU Junrui1, WU Rina1, YUE Xiqing1,*   

  1. (1. College of Food Science, Shenyang Agricultural University, Shenyang 110866, China; 2. College of Science, Liaoning Technology University, Fuxin 123000, China)
  • Online:2019-01-25 Published:2019-01-22

摘要: 采用杂交瘤技术从90株融合株中筛选得到分泌抗牛免疫球蛋白G(immunoglobulin G,IgG)单克隆抗体的2F6、5E3、9C6、9H8四个细胞株。其抗体亚型均为IgG1型,腹水单克隆抗体的效价可达5.12×106;5E3、9H8识别牛IgG重链Fc片段,2F6识别牛IgG轻链Fab片段,9C6识别牛IgG重链Fab片段;9C6与牛IgG1和牛IgG2反应,与山羊IgG、绵羊IgG、兔IgG、牛IgM、牛乳酪蛋白、牛乳β-乳球蛋白、牛乳铁蛋白、牛血清白蛋白、鸡卵清白蛋白、鱼明胶的交叉反应率均小于1%;9C6亲合力常数达牛乳8.92×108?L/mol。

关键词: 牛免疫球蛋白G(IgG), 单克隆抗体, 杂交瘤, 交叉反应, 亲合力

Abstract: Four hybridomas secreting monoclonal antibody (McAb) against bovine immunoglobulin G (IgG) were developed by fusion of spleen cells from immunized BALB/C mice with SP2/0 myeloma cells, and they were named as 2F6, 5E3, 9C6 and 9H8. The McAbs secreted by all four hybridomas were identified as IgG1 subtype with a titer of 5.12 × 106. Both 5E3 and 9H8 recognized the heavy chain Fc fragment of bovine IgG. 2F6 recognized the light chain Fab fragment of bovine IgG, and 9C6 recognized the heavy chain Fab fragment of bovine IgG. The McAb from 9C6 could react with both IgG1 and IgG2. The cross-reactivity between 9C6 and goat IgG, sheep IgG, rabbit IgG, bovine IgM, casein, β-lacto globulin, lactoferrin, bovine serum albumin, ovalbumin or fish gel was less than 1%. The affinity constant of 9C6 was 8.92 × 108 L/mol.

Key words: bovine immunoglobulin G, monoclonal antibody, hybridoma, cross-reactivity, affinity

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