| 초록 |
Development of effective antigens is demanded for efficient diagnosis and treatment of the parasite infection. While it is required to express and purify the recombinant proteins of the parasite for screening of novel antigens, cell-based expression method is time consuming and laborious. In this study, we explored a novel approach of cell-free expression and in situ immobilization of antigen to overcome the present limitations in antigen screening procedures. 5`-biotinylated PCR products of the genes were immobilized on agarose bead coated with streptavidin. When the DNA-conjugated agarose beads were incubated in a reaction mixture for cell-free synthesis, expressed proteins were immobilized on the same bead. Through this method, we were able to discover novel proteins that showed strong antigenicity. We expect that the effectiveness and throughput of cell-free expression and screening of antigenic proteins can be further improved when integrated with FACS. |
