[Role of molecular adjuvants TBhsp and MT in the preparation of monoclonal antibody against phosphatase of regenerating liver-3].

OBJECTIVE:To explore the adjuvant effects of Mycobacterium tuberculosis heat shock protein (TBhsp) and Mycobacterium tuberculosis T cell stimulatory epitope (MT) in the preparation of the monoclonal antibody (mAb) against phosphatase of regenerating liver-3 (PRL-3). METHODS:The prokaryotic expression vectors pET28a were used for the construction of pET28a-PRL-3 (control plasmids), pET28a-PRL-3-MT, pET28a-TBhsp-PRL-3 and pET28a- TBhsp-PRL-3-MT (recombinant plasmids). The various fusion proteins expressed in bacteria were purified and utilized to immunize the BALB/c mice, respectively. The serum level of specific anti-PRL-3 antibody was measured by ELISA. The mouse with the highest serum level of anti-PRL-3 antibody was selected to prepare mAb with hybridoma technique. The subclasses of mAb were identified. RESULTS:The PRL-3 fusion proteins expressed from the four expression plasmids were purified successfully. The serum level of anti-PRL-3 antibody from the mice immunized with PRL-3-MT was the highest compared with the other immune groups. Ten hybridoma cell lines secreting anti-PRL-3 mAb were obtained after cell fusion and ELISA primary screening, and all of the mAb subclasses were IgG. CONCLUSION:MT is a potentially effective molecular adjuvant in preparation of mAb specific for PRL-3.

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