Gene cloning, bacterial expression, in vitro refolding, and characterization of a single-chain Fv antibody against PreS1(21-47) fragment of HBsAg
文献类型：期刊论文
| 作者 | Yang, XX; Hu, WG; Li, F; Xia, HC; Zhang, ZC |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2005 |
| 卷号 | 41期号:2页码:341-348 |
| 关键词 | PreS1(21-47) HBsAg ScFv refolding expression gel filtration Escherichia coli |
| 通讯作者 | Zhang, ZC (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Key Lab Proteom, Inst Biochem & Cell Biol, Shanghai 200031, Peoples R China.,zhangzc@sunm.shcnc.ac.cn |
| 英文摘要 | The murine monoclonal antibody 125E11 is an IgG which recognizes PreS 1(21-47) fragment of large hepatitis B surface antigen. It has been successfully used for clinical detection of HBV virion in serum of hepatitis B patients. In present study, the genes of variable region in heavy chain (VH) and light chain (VL) of 125E11 have been cloned. Sequence analysis of cloned VH gene and VL gene showed that they had general characterization of immunoglobin variable region genes. According to Kabat classification, VH gene and VL gene belong to VH10 family, subgroup HID and V-K family subgroup 1, respectively. An expression vector of 125E11 single-chain Fv antibody fusion protein, in which VH and VL peptide were connected by a flexible linker (Gly(4)Ser)(3), was constructed. The scFv fusion protein was highly expressed in Escherichia coli mainly in inclusion body form. Using urea and pH gradient gel filtration method, the refolding of scFv was efficiently achieved. The refolding efficiency reached about 11 % and 2.7 mg refolded scFv was obtained from 1 L of culture. The binding activity and specificity of 125E11 scFv against PreS1 (21-47)-containing antigen were also analyzed. (c) 2005 Elsevier Inc. All rights reserved. |
| 学科主题 | Biochemistry & Molecular Biology; Biotechnology & Applied Microbiology |
| 类目[WOS] | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 关键词[WOS] | HEPATITIS-B-VIRUS ; SIZE-EXCLUSION CHROMATOGRAPHY ; ESCHERICHIA-COLI ; PROTEINS ; BINDING ; INFECTION ; DOMAINS |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000229192800013 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/1858]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Yang, XX,Hu, WG,Li, F,et al. Gene cloning, bacterial expression, in vitro refolding, and characterization of a single-chain Fv antibody against PreS1(21-47) fragment of HBsAg[J]. PROTEIN EXPRESSION AND PURIFICATION,2005,41(2):341-348. |
| APA | Yang, XX,Hu, WG,Li, F,Xia, HC,&Zhang, ZC.(2005).Gene cloning, bacterial expression, in vitro refolding, and characterization of a single-chain Fv antibody against PreS1(21-47) fragment of HBsAg.PROTEIN EXPRESSION AND PURIFICATION,41(2),341-348. |
| MLA | Yang, XX,et al."Gene cloning, bacterial expression, in vitro refolding, and characterization of a single-chain Fv antibody against PreS1(21-47) fragment of HBsAg".PROTEIN EXPRESSION AND PURIFICATION 41.2(2005):341-348. |
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