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可溶性HLA-G1-肽复合物的体外折叠和鉴定 被引量：2

Refolding and identification of soluble HLA-G1-peptide complex in vitro
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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.
作者 夏芳珍 吴雄文 钟茂华 褚利霞 翁秀芳 张彩娥 卢小玲 梁智辉
机构地区 华中科技大学同济医学院免疫学系
出处 《华中医学杂志》 CAS 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)。
关键词 基因 MHCI类 可溶性 HL-G1-肽复合物 原核表达 稀释复性 Gene, MHC class I Soluble HLA-G1-peptide oomplex Pmkaryotic expressing Refolding by dilution
分类号 R392 [医药卫生—免疫学]
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参考文献7

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同被引文献18

  • 1杨敬宁,吴雄文,梁智辉,陆盛军,尹姝晗,蔡蕾,韩军艳,黄亚非,龚非力.可溶性HLA-G1重链分子的构建、表达及纯化[J].中国免疫学杂志,2005,21(4):287-290. 被引量：4
  • 2陆盛军,梁智辉,朱慧芬,吴雄文,龚非力.抗HLA-G单克隆抗体G11E5的制备[J].细胞与分子免疫学杂志,2006,22(2):216-216. 被引量：1
  • 3Athanassakis I, Paflis M, Ranella A, et al. Detection of soluble HLA- G level in maternal serum can be predictive for a successful pregnancyl. Transplant Proc, 1999, 31(4): 1834-1837.
  • 4Casro MJ, Morales P, Rojo-Amigo R, et al. Homozygous HLA-G * 0105N healthy individuals indicate that membrane-associated HLA-G1 molecule is not necessary for survival. Tissue Antigens, 2000, 56(3) : 232-239.
  • 5Cre'put C, Durrbach A, Menier C, et al. Human leukocyte antigen-G (HLA-G) expression in biliary epithelial cells is associated with allograft acceptance in liver-kidney transplantation. J Hepatol, 2003, 39 (4) : 587-594.
  • 6Lila N, Amrein C, Guillemain R, et al. Human leukocyte antigen-G expression after heart transplantation is associated with a reduced incidence of rejection. Circulation, 20(0, 105(16): 1949-1954.
  • 7Lee N, Malacko AR, Ishitani A, et al. The membrane-bound and soluble forms of HLA-G bind identical sets of endogenous peptides but differ with respect to TAP association. Immunity, 1995, 3(5): 591-600.
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  • 9Lila N, Rouas-Freiss N, Dausset J, et al. Soluble HLA-G protein secreted by allo-specific CD4^+ T cells suppresses the allo-proliferative response: a CD4^+ T cell regulatory mechanism. Proc Natl Acad Sci USA, 2001, 98(21) : 12150-12155.
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华中医学杂志
2006年 第1期

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