可溶性HLA-G1-肽复合物的体外折叠和鉴定  

Refolding and identification of soluble HLA-G1-peptide complex in vitro

作　　者：夏芳珍[1] 吴雄文[1] 钟茂华[1] 褚利霞[1] 翁秀芳[1] 张彩娥[1] 卢小玲[1] 梁智辉[1] 

机构地区：[1]华中科技大学同济医学院免疫学系

出　　处：《华中医学杂志》2006年第1期7-9,共3页Central China Medical Journal

基　　金：国家自然科学基金资助项目(No.30271201);科技部973计划资助课题(No.2001CB510008)。

摘　　要：目的合成可溶性HLA-G1-肽复合物。方法利用原核高效表达的可溶性HLA-G1重链和轻链(β2m),在人工合成的HLA-G1限制性九肽(KGPPAALTL)存在的条件下,通过稀释复性折叠成可溶性HLA-G1-肽复合物。利用特异性抗体(W6/32及兔抗人β2m抗体)进行免疫印迹及酶联免疫吸附试验,检测稀释复性的折叠产物。结果折叠复合物含有35%可溶性HLA-G1-肽复合物、5%可溶性HLA-G1重链聚合体及60%复性的β2m3种成分。结论通过原核表达、体外折叠能够获得大量可溶性HLA-G1-肽复合物。Objective To refold soluble HLA-Gl-peptide complex in vitro. Methods The heavy chain of soluble HLA-G1 and β2m were high-level expressed as insoluble aggregates in E, ooli, and then the two subunits were refolded to form a sHL-A-G1-peptide complex by dilution method in the presence of a synthesized nonapeptide （KGPPAALTL）. The refolded product was detected by ELISA and Westem blot with mAb W6/32 and rabbit anti-humaβ2m antibody. Results The refo[ded complex was complesed of 35% sHLA-G1-peptide complex, 5% soluble heavy chain aggregate and 60% refolded β2m. Conclusion The refolding of sHLA G1 peptide complex can be successfully generated with Prokaryotie expressing and Refolding in vitro.

关 键 词：基因 MHCI类 可溶性 HL-G1-肽复合物 原核表达 稀释复性 

分 类 号：R392[医药卫生—免疫学]