Carcinogenesis, Teratogenesis & Mutagenesis

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Preparation and identification of monoclonal antibody against liver fatty acid binding protein

SONG Wei,YANG Hai-bo,CHEN Lan-ying*   

  1. College of Life Science and Engineering, Henan University of Urban Construction, Pingdingshan 467044, Henan, China
  • Received:2013-09-04 Revised:2014-01-24 Online:2014-05-30 Published:2014-05-30

Abstract:

OBJECTIVE: To prepare anti-liver-type fatty acid binding protein (LFABP) monoclonal antibody (mAb) and identified subtype and specificity. METHODS:The LFABP recombinant protein was used to immunize BALB/c mice,spleen cells from immunized mice were fused with myeloma cell Sp2/0 . After HAT selective culture and indirect ELISA screening,we obtained hybridoma cell line secreting mouse mAb against human LFABP. The specificity was verified by ELISA and Western blotting. RESULTS:After using purified recombinant protein immunized mice and screening,we obtained two hybridoma cell lines secreting the mAb against human LFABP,named 3E6 and 5B7,both were of the IgG1 subtype. The antibody was purified,reaching a concentration of 2 mg/mL,and titer of 1∶10 000 above. Western blotting showed that mAb had a specific reaction with the LFABP endogenous expression in the cells. CONCLUSION:We successfully prepared mice mAb against human LFABP ,which could not only provide an important foundation to further studies of the biological characteristics of LFABP,mechanism involved in regulation of fatty acid metabolism and drug interaction,but also provide experimental evidence for the treatment of related diseases.

Key words: liver-type fatty acid binding protein, prokaryotic expression, hybridoma, monoclonal antibody