[Improvement of refolding efficiency of HLA-A2-peptide complex in vitro by preformed disulfide bonds in heavy chain]

[Improvement of refolding efficiency of HLA-A2-peptide complex in vitro by preformed disulfide bonds in heavy chain] Research Abstract Details 

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[Improvement of refolding efficiency of HLA-A2-peptide complex in vitro by preformed disulfide bonds in heavy chain] Abstract Text:

Shi-jiang Fei,Xiong-wen Wu,Xiu-fang Wong,Lei Cai,Zhi-hui Liang,Jun-yan Han,Fei-li Gong,

AIM: To improve the refolding efficiency of soluble HLA-A2-peptide complex in vitro. METHODS: The heavy chain (HC) of MHC class I was extracted from bacteria under denaturing and non-reducing conditions. Anion-exchange and (NH4)2SO4 precipitation were applied to purify the HC. Then the purified HC, beta2m and an antigenic peptide (N-YMDGTMSQV-COOH of Try(369-377)) were refolded to form an HLA-A2-peptide complex by dilution method in the buffer of pH 6.6. The refolded products were detected by Western blot and ELISA with W6/32 and anti-human beta2m antibody. RESULTS: The refolded products consisted of HLA-A2-peptide complex, beta2m, and a little amount of HC polymer. The refolding efficiency was 2.5 fold higher than that of the conventional method. CONCLUSION: This study confirmed that the refolding efficiency of the method reported in this paper is higher as compared with the conventional method, which is of importance to the preparation of HLA-peptide tetramers and artificial antigen presenting cells.

[Improvement of refolding efficiency of HLA-A2-peptide complex in vitro by preformed disulfide bonds in heavy chain] Publishing Authors By Initials

SJ Fei,XW Wu,XF Wong,L Cai,ZH Liang,JY Han,FL Gong,

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[Improvement of refolding efficiency of HLA-A2-peptide complex in vitro by preformed disulfide bonds in heavy chain] Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Xi bao yu fen zi mian yi xue za zhi = Chinese jour

VOLUME: 21

Page Numbers: 167-70

Journal Abbreviation: Xi Bao Yu Fen Zi Mian Yi Xue Z

ISSN: 1007-8738

DAY: 15

MONTH: Mar

YEAR: 2005

[Improvement of refolding efficiency of HLA-A2-peptide complex in vitro by preformed disulfide bonds in heavy chain] Information

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LANGUAGE: chi

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AFFILIATION: Department of Immunology, Tongji Medical College, Huazhong University of Sciences and Technology, Wuhan 430030, China.

Country: China

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MEDLINETA: Xi Bao Yu Fen Zi Mian Yi Xue Z

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