| 用多糖-破伤风类毒素结合苗制备抗流行性脑膜炎球菌多糖单克隆抗体 |
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| 引用本文： | 李佩珊,张春燕,李时君,陈妍,项美娟,李方和,张洪. 用多糖-破伤风类毒素结合苗制备抗流行性脑膜炎球菌多糖单克隆抗体[J]. 医药导报, 2011, 30(6): 709-712. DOI: 10.3870/yydb.2011.06.005 |
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| 作者姓名： | 李佩珊 张春燕 李时君 陈妍 项美娟 李方和 张洪 |
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| 作者单位： | 1. 武汉大学第一临床学院药剂科,430060
2. 华中科技大学同济医学院附属同济医院实验医学研究中心,武汉,430030 |
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| 摘 要： | ［摘要］目的研制抗A群奈瑟脑膜炎球菌荚膜多糖单克隆抗体（GAMP mAb）。方法以A群奈瑟脑膜炎球菌荚膜多糖 破伤风类毒素（GAMP TT）耦联物免疫BALB/c小鼠，用常规细胞融合与克隆化技术获得一株能稳定分泌抗GAMP mAb的杂交瘤细胞株（2E7）。采用秋水仙素阻断法测定其染色体数目，降植烷诱导法制备含该抗体的小鼠腹腔积液，辛酸 硫酸铵沉淀法纯化单克隆抗体，十二烷基硫酸钠 聚丙烯酰胺凝胶电泳（SDS PAGE）灰度扫描测定提取物纯度，改良过碘酸盐氧化法制备辣根过氧化物酶（HRP）标记抗GAMP mAb，并采用中和法及其抗原替代法对抗GAMP mAb的特异性进行初步的鉴定。结果所获得的杂交瘤细胞（2E7）具有很好的生长特性，染色体数目约为139.73条；抗体分泌量适中，Ig亚类为IgG1,提取物浓度1.76～2.32 mg•mL 1，纯度97.2%，标记抗GAMP mAb的效价为1：25 600；抗原替代实验与中和实验对其特异性考核，初步结果证实2E7 mAb具有很好的抗GAMP特异性。结论成功制备抗GAMP mAb，为GAMP TT疫苗的生产监测及产品质量控制提供了必要的物质基础。
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| 关 键 词： | ［关键词］菌苗；多糖抗体；A群奈瑟脑膜炎球菌荚膜多糖；结合苗；单克隆抗体 |
Preparation of Monoclonal Antibody Against Neisseria Meningococcal Serogroup A Polysaccharide (GAMP) by Using the Conjugate Vaccine of both GAMP and Tetanus Toxoid |
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| LI Pei-shan,ZHANG Chun-yan,LI Shi-jun,CHEN yan,XIANG Mei-juan,LI Fang-he,ZHANG Hong. Preparation of Monoclonal Antibody Against Neisseria Meningococcal Serogroup A Polysaccharide (GAMP) by Using the Conjugate Vaccine of both GAMP and Tetanus Toxoid[J]. Herald of Medicine, 2011, 30(6): 709-712. DOI: 10.3870/yydb.2011.06.005 |
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| Authors: | LI Pei-shan ZHANG Chun-yan LI Shi-jun CHEN yan XIANG Mei-juan LI Fang-he ZHANG Hong |
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| Affiliation: | LI Pei-shan1,ZHANG Chun-yan2,LI Shi-jun1,CHEN yan2,XIANG Mei-juan1,LI Fang-he1,ZHANG Hong1(1.Department of Pharmacy,the First Clinical Medical Collegy,Wuhan University,Wuhan 430060,China,2.Central of Medicine,Tongji Hospital Affiliated with Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China) |
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| Abstract: | Objective To prepare monoclonal antibodies against Neisseria meningococcal serogroup A polysaccharide(GAMP). Methods BALB/c mouse was immunized by GAMP-Tetanus Toxoid(TT).Hybridoma cell lines secreting monoclonal antibodies against GAMP were screened after the fusion of mouse splenic cells with SP2/0 cells.The number of chromosome in hybridoma cell was tested by colchicines-interception,and the ascite of mice was prepared by pristane-inducing method.The mAb was purified by octansic acid-ammonium sulfate pre... |
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| Keywords: | Bacterial vaccine Polysaccharide antibody Neisseria meningococcal serogroup A polysaccharide Conjugate vaccine Monoclonal antibody |
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