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

AIM: To express HLA-A2 heavy chain (HC) and light chain β2m in bacteria and to prepare soluble HLA-A2-peptide complex. METHODS: HC-and β2m-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 chro-matography. Then HC and β2m 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 β2m 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℃ 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.