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编码针对乙型肝炎病毒前S2抗原的单克隆抗体重链和轻链的cDNA的克隆与鉴定

Cloning and characterization of cDNAs coding for heavy and light chains of a monoclonal antibody specific for pre-S2 antigen of hepatitis B virus.

作者信息

Hong H J, Kim A K, Ryu C J, Park S S, Chung H K, Kwon K S, Kim K L, Kim J, Han M H

机构信息

Genetic Engineering Research Institute, KIST, Daejeon, Korea.

出版信息

Gene. 1992 Nov 16;121(2):331-5. doi: 10.1016/0378-1119(92)90139-g.

Abstract

Binding specificity of a monoclonal antibody (mAb) (kappa, gamma 2b) H8 which can react with the pre-S2 peptide of hepatitis B virus (HBV) was determined by Western blot analyses. From the hybridoma cell line secreting mAb H8, poly(A)+ RNA was prepared and used as a template for cDNA synthesis and cloning. Full-length cDNAs coding for the heavy and kappa light chains of the mAb were cloned from the cDNA library and characterized by nucleotide (nt) sequence analyses and N-terminal amino acid sequencing. The sequence analyses revealed that both heavy and light chain-specific cDNAs are functional, and the variable regions of the heavy and light chains are members of mouse heavy chain subgroup III(c) and light chain group I, respectively. Comparison of the nt sequences with mouse immunoglobulin genes listed in the GenBank data base show that the cDNAs have not been previously reported. The cDNAs will be used for the construction of a therapeutic antibody for HBV infection.

摘要

通过蛋白质印迹分析确定了一种可与乙型肝炎病毒(HBV)前S2肽反应的单克隆抗体(mAb)(κ链,γ2b)H8的结合特异性。从分泌mAb H8的杂交瘤细胞系中制备了聚腺苷酸加尾(poly(A)+)RNA,并将其用作cDNA合成和克隆的模板。从cDNA文库中克隆出编码该mAb重链和κ轻链的全长cDNA,并通过核苷酸(nt)序列分析和N端氨基酸测序进行表征。序列分析表明,重链和轻链特异性cDNA均具有功能,重链和轻链的可变区分别属于小鼠重链III(c)亚组和轻链I组。将nt序列与GenBank数据库中列出的小鼠免疫球蛋白基因进行比较,结果表明这些cDNA此前尚未见报道。这些cDNA将用于构建针对HBV感染的治疗性抗体。

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