[Preparation and preliminary characterization of soluble HLA-A2-peptide complex].
OBJECTIVE
To express HLA-A2 heavy chain (HC) and light chain beta(2m) in bacteria and to prepare soluble HLA-A2-peptide complex.


METHODS
HC and beta(2m) expressing engineering bacteria was induced for 5 h with 0.5 mmol/L IPTG. After sonification of the bacteria,crude inclusion body was obtained which was then refined, renaturated, ultrafiltrated, and purified by DEAE Sepherose Fast Flow anion exchange chromatography. Then HC and beta(2m) were connected with two specific peptides, purified through Superdex 75 gel filtration, and identified with mAb W6/32 which can recognize native HLA-A2.


RESULTS
The expression rates of HC and beta(2m) in engineering bacteria was both about 50%. The purity of both expression products reached to 95%.Moreover, the two HLA-A2-peptide complexes could be recognized by mAb W6/32, even after being stored at 4 degrees Celsius for 2 months.


CONCLUSIONS
The two soluble HLA-A2-peptide complexes prepared by us are stable and lay the foundation for further research of CTL recognition and response.
Meng
Xiao-jing
XJ
Institute of Molecular Immunology, First Military Medical University, Guangzhou 510515, China.
Linlai
Xing-mei
XM
Meng
Min-jie
MJ
Hu
Zhi-ming
ZM
Wang
Xiao-ning
XN
D015789:HLA-A2 Antigen / Q000378:metabolism*; D007218:Industrial Microbiology; D010455:Peptides / Q000378:metabolism*; D012995:Solubility; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D001613:beta 2-Microglobulin / Q000378:metabolism
D015789:HLA-A2 Antigen; D010455:Peptides; D001613:beta 2-Microglobulin