Preparation of monoclonal antibodies against pneumococcal polysaccharide and hepatitis B virus surface protein

Q4 Immunology and Microbiology 中华微生物学和免疫学杂志 Pub Date : 2019-12-31 DOI:10.3760/CMA.J.ISSN.0254-5101.2019.12.008
Qian Wen, Ying Zhang, Chen Nanping, Chen Yuqiu, W. Lili, Kai Wu, C. Min, Shi Jing, Qihan Li
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Abstract

Objective To prepare monoclonal antibodies against pneumonia serotype 33F polysaccharides (Pn33Fps) and hepatitis B virus (HBV) surface proteins (HBs) by using the conjugate of Pn33Fps and HBs as antigen. Methods The conjugate of Pn33Fps and HBs was used as antigen to immunize mice with different immune doses, different immune procedures and different immune sites. Mouse spleen cells with higher antibody level after immunization were isolated and fused with SP2/0 myeloma cells. The hybridoma cells were screened specifically with Pn33Fps or HBs to prepare corresponding monoclonal antibodies. Results Serum antibodies against Pn33Fps and HBs were induced by immunizing mice with the conjugate. Monoclonal cell lines capable of continuously expressing antibodies against Pn33Fps or HBs were obtained. It has been proved that the recovery rates of samples of Pn33Fps and HBs prepared in three batches tested with ascites monoclonal antibodies prepared by these two monoclonal cell lines were between 95% and 105%. Conclusions Monoclonal antibodies against Pn33Fps and HBs could be prepared simultaneously by immunizing mice with the conjugate of Pn33Fps and HBs and used for the quantitative detection of Pn33Fps and HBs. Key words: Conjugates; Monoclonal antibody; Pneumonia polysaccharide; Hybridoma cells; Hepatitis B virus surface protein
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抗肺炎球菌多糖和乙型肝炎病毒表面蛋白单克隆抗体的制备
目的以肺炎血清型33F多糖(Pn33Fps)和乙型肝炎病毒(HBV)表面蛋白(HBs)偶联物为抗原制备抗肺炎血清型33F多糖(Pn33Fps)和乙型肝炎病毒(HBV)表面蛋白(HBs)的单克隆抗体。方法采用Pn33Fps与HBs结合物作为抗原,对不同免疫剂量、不同免疫程序和不同免疫部位的小鼠进行免疫。分离免疫后抗体水平较高的小鼠脾细胞,与SP2/0骨髓瘤细胞融合。用Pn33Fps或HBs特异性筛选杂交瘤细胞,制备相应的单克隆抗体。结果用该偶联物免疫小鼠可诱导血清抗Pn33Fps和HBs抗体。获得了能够连续表达Pn33Fps或HBs抗体的单克隆细胞系。实验证明,用这两种单克隆细胞系制备的腹水单克隆抗体检测三批制备的Pn33Fps和HBs样品的回收率在95% ~ 105%之间。结论将Pn33Fps和HBs结合物免疫小鼠,可同时制备抗Pn33Fps和HBs的单克隆抗体,用于Pn33Fps和HBs的定量检测。关键词:共轭;单克隆抗体;肺炎多糖;杂种瘤细胞;乙型肝炎病毒表面蛋白
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来源期刊
中华微生物学和免疫学杂志
中华微生物学和免疫学杂志 Immunology and Microbiology-Virology
CiteScore
0.50
自引率
0.00%
发文量
6906
期刊介绍: Chinese Journal of Microbiology and Immunology established in 1981. It is one of the series of journal sponsored by Chinese Medical Association. The aim of this journal is to spread and exchange the scientific achievements and practical experience in order to promote the development of medical microbiology and immunology. Its main contents comprise academic thesis, brief reports, reviews, summaries, news of meetings, book reviews and trends of home and abroad in this field. The distinguishing feature of the journal is to give the priority to the reports on the research of basic theory, and take account of the reports on clinical and practical skills.
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