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人源抗NH-LBP Fab抗体的制备和初步鉴定 被引量：2

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摘要 目的构建人源抗脂多糖结合蛋白氨基端片段(NH-LBP)Fab抗体的原核表达载体,并制备高稳定性Fab抗体。方法用PCR方法,从可溶性表达抗NH-LBP抗体的重组质粒pComb3H-Fab中扩增出VH链和VL链基因序列,分别构建表达质粒pET-28a(+)-VH和pET-28a(+)-VL,并于工程菌BL21(DE3)star中分别表达及TALON柱芯亲和纯化,将抗体片段VH和VL共同复性,通过二硫键形成Fab抗体,再采用ELISA法初步鉴定其与NH-LBP的结合力。结果扩增出VL链和VH基因片段长约为650bp和700bp,经BL21star表达出相对分子质量(Mr)约为28000和30000的目的蛋白,共同折叠后所获得Fab抗体与NH-LBP具有较高的结合力。结论成功地制备抗NH-LBPFab抗体,为临床抗炎研究提供了条件。
作者 葛晓冬 邹佳 刘友生 杨艳丽 段光杰
机构地区 第三军医大学第一附属医院病理科
出处 《细胞与分子免疫学杂志》 CAS CSCD 北大核心 2007年第4期359-362,共4页 Chinese Journal of Cellular and Molecular Immunology
基金 国家自然科学基金资助项目(30400426) 第三军医大学校科研基金(2004年)
关键词 脂多糖结合蛋白氨基端片段(NH-LBP) 人源抗体Fab 高效表达
分类号 R392.11 [医药卫生—免疫学]
  • 相关文献

参考文献10

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二级参考文献7

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共引文献10

同被引文献18

  • 1葛晓冬,刘友生,王晓东,王长松,邓军.重组人脂多糖结合蛋白的氨基末端片段在昆虫细胞中的表达、纯化及功能鉴定[J].第三军医大学学报,2004,26(22):2029-2032. 被引量：1
  • 2葛晓冬,邹佳,杨艳丽,刘友生.抗人脂多糖结合蛋白氨基端片段抗体dsFv V_L链的构建、表达和鉴定[J].第三军医大学学报,2007,29(14):1364-1367. 被引量：1
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引证文献2

二级引证文献1

细胞与分子免疫学杂志
2007年 第4期

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