[Preparation and preliminary characterization of soluble HLA-A2-peptide complex].

ABSTRACT

AIM: 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. CONCLUSION: The two soluble HLA-A2-peptide complexes prepared by us are stable and lay the foundation for further research of CTL recognition and response.