Integrative Materials Research Institute, Hallym University, Chuncheon-si, Republic of Korea; Department of Materials Science and Engineering, Hallym University, Chuncheon-si, Republic of Korea.
Department of Materials Science and Engineering, Yonsei University, Seoul, Republic of Korea.
J Pharm Biomed Anal. 2018 Jan 5;147:174-184. doi: 10.1016/j.jpba.2017.07.043. Epub 2017 Jul 31.
Autodisplay is an expression system for the display of recombinant proteins on the outer membrane (OM) of gram negative bacteria and has been developed for translocation studies, whole cell biocatalysis, bioremediation, inhibitor screening, and enzyme refolding. Recently, affinity proteins such as IgG-binding Z-domains and biotin-binding streptavidin have been autodisplayed on the OM of Escherichia coli for analytical and biomedical applications. The secretion mechanism of the autodisplay system was used and orientation and density control of these affinity proteins were determined. Affinity protein-autodisplaying E. coli cells have been used to coat solid supports in immunoassays. For this purpose, the OM of autodisplayed E. coli cells was separated and isolated by the aid of detergents. The structure of the resulting OM liposomes as well as their physico-chemical parameters, were analyzed. OM liposomes were used subsequently for coating various solid matters including microplates and biosensor transducer surfaces and the formation of OM layers were monitored. OM layer formation on solid matters was shown to increase the sensitivity of immunoassays and biosensors. In this review, analytical and diagnostic applications are described in particular concerning orientation and density control of autodisplayed affinity proteins.
自动展示是一种在革兰氏阴性菌外膜（OM）上展示重组蛋白的表达系统，已被开发用于转运研究、全细胞生物催化、生物修复、抑制剂筛选和酶重折叠。最近，免疫球蛋白结合 Z 结构域和生物素结合链霉亲和素等亲和蛋白已在大肠杆菌 OM 上自动展示，用于分析和生物医学应用。利用自动展示系统的分泌机制，确定了这些亲和蛋白的取向和密度控制。亲和蛋白自动展示大肠杆菌细胞已被用于免疫测定中的固体载体涂层。为此，借助去污剂分离和分离自动展示大肠杆菌细胞的 OM。分析了由此产生的 OM 脂质体的结构及其物理化学参数。随后，将 OM 脂质体用于涂覆各种固体物质，包括微孔板和生物传感器换能器表面，并监测 OM 层的形成。结果表明，OM 层在固体物质上的形成增加了免疫测定和生物传感器的灵敏度。在这篇综述中，特别描述了分析和诊断应用，涉及自动展示亲和蛋白的取向和密度控制。
