| PRRSV GP5蛋白抗独特型单链抗体的构建表达及鉴定 |
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| 引用本文： | 于颖,王刚,宋腾飞,孔宁,马晓春,姚永红,肖一红,周恩民.PRRSV GP5蛋白抗独特型单链抗体的构建表达及鉴定[J].中国预防兽医学报,2011,33(6). |
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| 作者姓名： | 于颖 王刚 宋腾飞 孔宁 马晓春 姚永红 肖一红 周恩民 |
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| 作者单位： | 1. 山东农业大学动物医学院,山东泰安,271018
2. 山东农业大学动物医学院,山东泰安271018;西北农林科技大学动物医学院,陕西杨凌712100 |
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| 基金项目： | 国家行业科技计划项目，国家自然科学基金，山东省自主创新成果转化重大专项 |
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| 摘 要： | 为制备只包含抗体可变区片段(Fv)的MAb2-5G2单链抗体(scFv),本研究从分泌MAb2-5G2的杂交瘤细胞中提取总RNA,反转录制备cDNA作为模板扩增重链可变区(VH)和轻链可变区(VL)基因.以VH-1inker-VL的形式通过重叠PCR制备scFv基因,构建重组表达质粒pEasy-scFv,并在大肠杆菌中获得正确表达.通过western blot和间接免疫荧光鉴定表明scFv蛋白能够被兔抗MAb2-5G2的抗体(Ab3)识别,复性后的scFv蛋白能够特异性结合Marc-145细胞.该研究结果为进一步明确MAb2-5G2结合Marc-145细胞功能区域奠定了物质基础.
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| 关 键 词： | 猪繁殖与呼吸综合征 抗独特型抗体 单链抗体 |
Construction, expression and identification of anti-idiotype scFv specific to anti-PRRSV GP5 antibody |
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| YU Ying,WANG Gang,SONG Teng-fei,KONG Ning,MA Xiao-chun,YAO Yong-hong,XIAO Yi-hong,ZHOU En-min.Construction, expression and identification of anti-idiotype scFv specific to anti-PRRSV GP5 antibody[J].Chinese Journal of Preventive Veterinary Medicine,2011,33(6). |
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| Authors: | YU Ying WANG Gang SONG Teng-fei KONG Ning MA Xiao-chun YAO Yong-hong XIAO Yi-hong ZHOU En-min |
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| Institution: | YU Ying1,WANG Gang1,SONG Teng-fei1,KONG Ning1,MA Xiao-chun1,YAO Yong-hong1,XIAO Yi-hong1*,ZHOU En-min1,2*(1.College of Animal Medicine,Shandong Agricultural University,Shandong 271018,China,2.College of Animal Medicine,Northwest A&F University,Yangling,712100,China) |
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| Abstract: | Anti-idiotype plays important roles in the immune responses against porcine reproductive and respiratory syndrome virus(PRRSV).Our previous study has shown that the monoclonal anti-idiotype antibody(MAb2) 5G2 specific to anti-PRRSV GP5 antibody could recognize and bind to Marc-145 cells.To further understand the functional domain of MAb2-5G2 for binging to Marc-145 cells,the variable light(VL) and variable heavy(VH) chain genes of MAb2-5G2 were amplified from the hybridoma cells secreting MAb2-5G2 by RT-PCR... |
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| Keywords: | porcine reproductive and respiratory syndrome virus anti-idiotype antibody scFv |
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