抗人氨基末端脂多糖结合蛋白二硫键稳定性Fv抗体的制备及生物学活性的初步研究  

Preparation and characterization of disulfide stabilized Fv fragment antibody against N-terminal fragment of human lipopolysaccharide binding protein

作　　者：邹佳[1] 葛晓冬[1] 杨艳丽[1] 刘友生[1] 

机构地区：[1]第三军医大学西南医院病理学研究所,重庆400038

出　　处：《第三军医大学学报》2008年第14期1308-1311,共4页Journal of Third Military Medical University

基　　金：国家自然科学基金(30400426);第三军医大学科研创新基金(2004)~~

摘　　要：目的制备抗人氨基末端脂多糖结合蛋白(N-terminal fragment of human lipopolysaccharide binding protein,NH-LBP)的二硫键稳定性Fv抗体(disulfide stabilized Fv fragment,dsFv),并初步测定其生物学活性。方法将含有dsFv-VL与dsFvVH包涵体蛋白经复性、折叠和纯化,获得完整的dsFv抗体。通过ELISA、TNF-α分泌等方法,初步测定dsFv抗体在体内外的生物学活性。结果获得dsFv抗体蛋白约2.1mg,dsFv与NH-LBP有较好的结合能力,并在动物体内减轻了LPS所诱导的炎症反应,发挥了一定的保护活性。结论通过分别表达VL和VH片段,再制备dsFv抗体是可行的,dsFv能部分抑制LBP的生物学功能。Objective To obtain the disulfide stabilized Fv fragment （dsFv） against N-terminal fragment of human lip polysaccharide binding protein （NH-LBP） and to identify its biological vitality. Methods The disulfide stabilized Fv fragment antibody （dsFv） was obtained after the inclusion bodies of dsFvVs and dsFvVr had been refolded and purified. Then the characteristics of dsFv were determined in vitro by ELISA and by detecting the secretion of tumor necrosis factor alpha （TNF-α） in rats. Results There was 2.1 mg protein of dsFv obtained, dsFv had good combination with NH-LBP and could restrain inflammatory reaction caused by lip polysaccharide （LPS） in vivo. Conclusion It is feasible to get dsFv against NH-LBP by respective expression of Vr and Vs. The partial inhibition of the biological function of LBP by dsFv is a new way to restrain the over-inflammatory reaction in vivo.

关 键 词：人氨基末端脂多糖结合蛋白 包涵体 二硫键稳定性Fv抗体 

分 类 号：R392-33[医药卫生—免疫学] R392.11[医药卫生—基础医学]