Preparation of monoclonal antibodies against Mycobacterium tuberculosis TB10.4 antigen.

TB10.4 protein is a member of the ESX family that is necessary for Mycobacterium tuberculosis survival and plays a vital role in mycobacterial pathogenesis. In this study, the gene encoding TB10.4 was cloned into prokaryotic expression vecters pET-30(a) and pGEX-6p-1. The two recombinant proteins His-TB10.4 and GST-TB10.4 were then expressed in vitro in prokaryotic expression systems to develop monoclonal antibodies (MAbs) against TB10.4 protein. The purified rHis-TB10.4 protein was used to immunize BALB/c mice, and eight MAbs were produced. An immunoblotting analysis indicated that all these MAbs specifically recognize the TB10.4 protein. These new MAbs provide powerful reagents for further functional research into TB10.4 protein.