Assay development for analyzing high performance antibody selection based on cell-free synthesized GPCR proteoliposome.

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We developed a high-throughput screening method using cell-free synthesized proteoliposome based on AlphaScreen, ELISA, Biacore, respectively. AlphaScreen and ELISA had excellent throughput, thus suitable for primary screening of monoclonal antibodies. Especially AlphaScreen was superior for conformational sensitive antibody selection. Using the methods developed in this study, we conducted screening and functional analysis of anti-GPCR monoclonal antibody. We immunized mice with cell-free synthesized and purified DRD1 proteoliposome and developed 800 hybridoma lines. Primary screening of 800 hybridoma was performed using AlphaScreen and ELISA, respectively, and 36 specific monoclonal antibodies were selected. They were high affinity and applicable for multiple applications. These results demonstrated that the technology based on cell-free synthesized GPCR proteoliposome may provide advantageous materials and assay methods for monoclonal antibody production.