小鼠TLR2胞外区的克隆表达及单克隆抗体的制备

【Objectives】: In order to provide tools for studying the function of TLR2 receptor in tumor metastases, the monoclonal antibodies against the extracellular domain of mouse TLR2 were prepared. 【Methods】: The extracellular domain of mouse TLR2 was cloned into the prokaryotic expression vector pGEX-4T-1. Then the E. coli strain BL21 was transformed with the recombinant plasmid pGEX-TLR2-ECD and the fusion protein TLR2-ECD-GST was highly expressed by IPTG induction. Bal b/c mice were immuned with purified recombinant protein TLR2-ECD-GST, and the hybridoma was prepared to produce monoclonal antibodies against TLR2-ECD. Following negative screening and positive selection steps by indirect ELISA, hybridoma cell strains with the ability to secrete specific antibodies against TLR2-ECD were identified. 【Results】: Hybridoma cell strains secreting monoclonal antibody against TLR2-ECD were prepared. 【Conclusions】: Hybridoma cell strains secreting monoclonal antibody specifically recognized by TLR2 extra cellular domain were established. No cross-reactivity of the antibodies with the GST tagged protein was observed.