ABSTRACT

Antibody discovery against complex antigens is limited by the availability of a reproducible pure source of concentrated properly folded antigen. We have developed a technology to enable direct incorporation of membrane proteins such as GPCRs and into the membrane of poxvirus. The protein of interest is correctly folded and expressed in the cell-derived viral membrane and does not require any detergents or refolding before downstream use. The poxvirus is selective in which proteins are incorporated into the viral membrane, making the antigen poxvirus an antigenically cleaner target for in vitro panning. Antigen-expressing virus can be readily purified at scale and used for antibody selection using any in vitro display platform.

摘要

针对复杂抗原的抗体发现受到浓缩正确折叠抗原的可重复纯源的可用性的限制。我们开发了一种技术，可以将 GPCR 等膜蛋白直接掺入痘病毒膜中。目的蛋白在细胞来源的病毒膜中正确折叠和表达，并且在下游使用之前不需要任何去污剂或重折叠。痘病毒具有选择性，其中蛋白质被整合到病毒膜中，使得抗原痘病毒成为体外淘选的抗原清洁靶标。表达抗原的病毒可以很容易地大规模纯化，并使用任何体外展示平台用于抗体选择。