| THE CONSTRUCTION AND EXPRESSION OF THE MURINE SCFV GENE IN E. COLI AGAINST HUMAN CERVICAL CANCER |
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| 作者姓名： | 王莹 陈葳 李旭 |
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| 作者单位： | Center for Laboratory Medicine, the First Affiliated Hospital of Medical School, Xi＇an JiaotongUniversity, Xi＇an 710061, China. |
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| 基金项目： | FoundationItem： This study was supported by the Foundation of Science ＆ Technique Investigation Project of Shaanxi Province （No. 99SM52）. |
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| 摘 要： | Objective To obtain the gene of murine Single chain Fv fragment （ScFv） against haman cervical cancer and to express it in E. coli. Methods The variable region gene fragments of the heavy and light chains, which were amplified respectively using recombinant DNA techniques from CsA125 hybridama cells, were spliced together through a flexible linker to ScFv against human cervical cancer. The ScFv genes were then cloned into expression vector pCANTAB 5E and expressed in E. coli HB2151 and TG1 respectively. The soluble ScFv were characterized by SDSPAGE and Western blot. The antigen-binding activities of the soluble and phage displayed ScFv were assayed by ELISA and cell immunohistochemical analysis. Results The expressed ScFv antibodies were soluble and phage displayed. soluble ScFv secreted and expressed in E. coli HB2151 induced by IPTG were confirmed with SDS-PAGE, Western blot and ELISA. The specific binding capacity of the soluble and phage displayed ScFv to the surface associated antigen of human cervical cancer cell line was further confirmed with immunohistochemical studies. Conclusion The soluble and phage displayed ScFv expressed in E. coll against haman cervical cancer showed high, specific affinity for the cervical cancer cell line surface associated antigen.
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| 关 键 词： | 基因表达 大肠杆菌 宫颈癌 病理机制 |
| 文章编号： | 1671-8267（2006）01-0053-04 |
THE CONSTRUCTION AND EXPRESSION OF THE MURINE SCFV GENE IN E. COLI AGAINST HUMAN CERVICAL CANCER |
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| Wang Ying,Chen Wei,Li Xu.THE CONSTRUCTION AND EXPRESSION OF THE MURINE SCFV GENE IN E. COLI AGAINST HUMAN CERVICAL CANCER[J].Academic Journal of Xi’an Jiaotong University,2006,18(1):53-56,93. |
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| Authors: | Wang Ying Chen Wei Li Xu |
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| Abstract: | Objective To obtain the gene of murine Single chain Fv fragment (ScFv) against human cervical cancer and to express it in E.coli . Methods The variable region gene fragments of the heavy and light chains, which were amplified respectively using recombinant DNA techniques from CsA125 hybridoma cells, were spliced together through a flexible linker to ScFv against human cervical cancer. The ScFv genes were then cloned into expression vector pCANTAB 5E and expressed in E.coli HB2151 and TG1 respectively. The soluble ScFv were characterized by SDS-PAGE and Western blot. The antigen-binding activities of the soluble and phage displayed ScFv were assayed by ELISA and cell immunohistochemical analysis. Results The expressed ScFv antibodies were soluble and phage displayed. The soluble ScFv secreted and expressed in E.coli HB2151 induced by IPTG were confirmed with SDS-PAGE, Western blot and ELISA. The specific binding capacity of the soluble and phage displayed ScFv to the surface associated antigen of human cervical cancer cell line was further confirmed with immunohistochemical studies. Conclusion The soluble and phage displayed ScFv expressed in E.coli against human cervical cancer showed high, specific affinity for the cervical cancer cell line surface associated antigen. |
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| Keywords: | cervical cancer Single chain Fv fragment (ScFv) phage displayed scFv |
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