可溶性HLA-G-肽复合物的体外制备与纯化  

Preparation and Purification of Soluble HLA-G-peptide Complex in vitro

作　　者：钟茂华[1] 翁秀芳[1] 梁智辉[1] 夏芳珍[1] 李佳楠[1] 陈雪玲[1] 吴雄文[1] 

机构地区：[1]华中科技大学同济医学院基础医学院免疫学系,武汉430030

出　　处：《华中科技大学学报（医学版）》2008年第3期377-379,共3页Acta Medicinae Universitatis Scientiae et Technologiae Huazhong

基　　金：国家自然科学基金(No30772040);国家重点基础研究发展规划(973)(No2007CB512900)资助项目

摘　　要：目的探讨可溶性HLA-G-肽复合物的体外制备与纯化。方法原核高效表达的HLA-G重链及轻链(β2m)在抗原肽(人工合成九肽NH2-KGPPAALTL-COOH)的存在下,通过稀释复性折叠成HLA-G-肽复合物,采用凝胶过滤层析对折叠复合物进行纯化。利用特异性抗体(mAb W6/32和兔抗人β2m抗体)进行ELISA和Western blot对纯化产物进行鉴定。结果折叠复合物中,主要含有HLA-G重链聚合体、HLA-G-肽复合物及β2m3种成分,纯化后主要为HLA-G-肽复合物。结论成功地获得纯度较高的可溶性HLA-G-肽复合物单体,为进一步研究HLA-G的生物学功能奠定了基础。Objective To prepare and purify soluble HLA-G-peptide complex in vitro. Methods The heavy chain of HLA- G1 and β2m were expressed highly as insoluble aggregates in E. coli, and then the two subunits were refolded to form an HLA- G-peptide complex by dilution method in the presence of an antigenic peptide （NH2-KGPPAALTL-COOH） and purified by gel laminar analysis. The purified products were detected by ELISA and Western blot with mAb W6/32 and rabbit antl-human β2m antibody. Results The refolded complex was composed of H chain aggregates, HLA-G-peptide complex and β2m. The purified product was mainly HLA-G-peptide complex. Conclusion The refolded HLA-G-peptide complex was successfully purified and the products were confirmed by the practical immunological method. This study lald the foundation for the preparation of HLA-G-peptide complex for the further study on HLA-G.

关 键 词：HLA-G-肽复合物 稀释复性 凝胶过滤层析 

分 类 号：R392.12[医药卫生—免疫学]