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钟彦伟, 成军, 王刚, 田小军, 陈新华, 李莉, 陈菊梅, 张玲霞. 乙肝病毒核心抗原人源单链可变区抗体的筛选[J]. 中国公共卫生, 2002, 18(2): 153-154. DOI: 10.11847/zgggws2002-18-02-15
引用本文: 钟彦伟, 成军, 王刚, 田小军, 陈新华, 李莉, 陈菊梅, 张玲霞. 乙肝病毒核心抗原人源单链可变区抗体的筛选[J]. 中国公共卫生, 2002, 18(2): 153-154. DOI: 10.11847/zgggws2002-18-02-15
ZHONG Yan-wei, CHENG Jun, WANG Gang, . Screening on Human Phage Antibody of Hepatitis B Virus Core Antigen[J]. Chinese Journal of Public Health, 2002, 18(2): 153-154. DOI: 10.11847/zgggws2002-18-02-15
Citation: ZHONG Yan-wei, CHENG Jun, WANG Gang, . Screening on Human Phage Antibody of Hepatitis B Virus Core Antigen[J]. Chinese Journal of Public Health, 2002, 18(2): 153-154. DOI: 10.11847/zgggws2002-18-02-15

乙肝病毒核心抗原人源单链可变区抗体的筛选

Screening on Human Phage Antibody of Hepatitis B Virus Core Antigen

  • 摘要: 目的 筛选、鉴定乙型肝炎病毒(HBV)核心抗原(HBcAg)蛋白的人源单链可变区抗体(ScFv)的编码基因,为细胞内表达小分子单链抗体的研究及抗HBV的基因治疗研究奠定基础。方法 采用噬菌体表面展示技术,以HBcAg蛋白为固相抗原,从噬菌体单链可变区抗体半合成库中经过5轮“吸附-洗脱-扩增”筛选过程,获得抗原结合活性较强的HBcAg人源单链可变区抗体阳性克隆,并对其进行免疫检测及序列测定。结果 筛选得到的ScFv片段编码基因为771nt,编码的产物由257个氨基酸残基组成,具有典型的轻链和重链可变区结构特点以及与HBcAg结合的特异性。结论 利用噬菌体抗体库技术成功地获得了HBcAg人源单链可变区抗体的编码基因,并获得了可溶性单链抗体的表达。

     

    Abstract: Objective To screen and characterize human-phage antibody of core antigen of hepatiti B virus(HBcAg).Methods Using HBc antigen as the coating antigen,the recombinant phage antibody to HBc antigen were panned from a semisyn-thetic phage library by phage display technique,after five rounds of biopanning,60 clones were demonstrated specific to HBcAg antigen.The specificity of ScFv was demonstrated by EL ISA.Results HBcAg phage antibody has a specific combinating capacity with HBcAg antigen.The data DNA sequence showed that the ScFv gene of HBcAg was composed of 771 bp.Conclusion Human-single-chain antibody of HBcAg antigen was identified by means of the phage display technique.

     

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