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可溶性HLA-G-肽复合物的体外制备与纯化 被引量：1

Preparation and Purification of Soluble HLA-G-peptide Complex in vitro
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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.
作者 钟茂华 翁秀芳 梁智辉 夏芳珍 李佳楠 陈雪玲 吴雄文
机构地区 华中科技大学同济医学院基础医学院免疫学系
出处 《华中科技大学学报（医学版）》 CAS CSCD 北大核心 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
关键词 HLA-G-肽复合物 稀释复性 凝胶过滤层析 HLA-G peptide complex dilution refolding gel laminar analysis
分类号 R392.12 [医药卫生—免疫学]
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参考文献10

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共引文献8

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华中科技大学学报（医学版）
2008年 第3期

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