Cloning and expression of mouse antiª²HBVª²TP monoclonal antibody light chain and heavy chain variable gene fragments

    GAO Ping1, WAN Xiaoª²Rong2, LIN Fang3, ZHANG Huiª²Zhong3

    1Department of Obstetrics & Gynecology, 3Center of Clinical Research, Tangdu Hospital, Fourth Military Medical University, Xi¬ðan 710038, China, 2No. 94370 Unit of Chinese PLA, Jinan 250023, China

    ¡¾Abstract¡¿ AIM: To amplify and express the light and heavy chain variable gene fragments of mouse antiª²HBV Terminal protein (HBVª²PT) monoclonal antibody. METHODS: Light and heavy chain variable gene fragments of mouse antiª²HBVª²PT monoclonal antibody were amplified by RTª²PCR using cell total RNA purified from antibody producing hybridoma cells (E3), and further cloned into pGEMª²T vector, which were then sequenced. The correct fragments were then inserted into PBV220 plasmid to construct prokaryotic expression vector. The inserts were expressed under 42¡æ condition and the expression products were identified by Western blot. RESULTS: E3Amb light and heavy chain gene fragments consisted of 363 bp and 375 bp of nucleotides encoding 121 and 125 amino acid residues respectively. The amino acid sequences of both variable regions were in agreement with the characterization of the amino acid present in the mouse Ig variable region. The Western blot result further confirmed these expression products as mouse antibody origin. CONCLUSION: The light and heavy chain variable gene fragments of mouse antiª²HBVª²PT antibody are successfully cloned and expressed. These variable gene fragments of the antibody can be used for further immunotherapy studies of HBV infected patients. ......