Prediction of HLAª²A2 restricted cytotoxic T lymphocyte epitopes of TAG1 and expression and purification of fusion proteins of these epitopes and Hsp70

    LUAN Qi, GAO Tianª²Wen, LI Chunª²Ying, WANG Gang, SHEN Zhu, DANG Yuª²Ping, LI Qiang

    Department of Dermatology, Xijing Hospital, Fourth Military Medical University, Xi¬ðan 710033, China

    ¡¾Abstract¡¿ AIM: To predict HLAª²A2 restricted cytotoxic T lymphocyte (CTL) epitopes of the novel cancer/testis antigen TAG1 and to express and purify the fusion proteins of these epitopes and Hsp70 in prokaryotic system. METHODS: Longª²distance prediction system SYFPEITHI and polynomial method were used to predict the HLAª²A2 restricted CTL epitopes of TAG1. The epitopes were then cloned into the expression vector after synthesizing and annealing the nucleotide sequences of these epitopes. After induced with IPTG, the expressed fusion proteins were purified with GSTrap FF column. RESULTS: Three HLAª²A2 restricted CTL epitopes of TAG1 were selected. The fusion genes PGEXª²bwª²Hsp70 were constructed and the fusion proteins were acquired successfully. CONCLUSION: The fusion proteins of Hsp70 and three HLAª²A2 restricted CTL epitopes are expressed and purified successfully, which will be helpful in studying their immune functions and develop new peptide vaccines of malignant melanoma. ......