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Info. Vol.7 - No.2 (2013.06.20)
Title Optimization of a FACS based-immunoassay using E. coli autodisplaying Z-domains
Authors Min Park1, Ji-Hong Bong1, Gu Yoo1, Joachim Jose2, Min-Jung Kang3 & Jae-Chul Pyun1,*
Institutions 1Department of Materials Science and Engineering, Yonsei University 50 Yonsei-Ro, Seodaemun-Gu, Seoul 120-742, Korea
2Institute of Pharmaceutical and Medical Chemistry, University of Muenster Muenster, Germany
3Korea Institute of Science and Technology (KIST), Seoul, Korea
Abstract We developed a fluorescence-activated cell sorter (FACS)-based immunoassay using Escherichia coli with autodisplayed Z-domains as a solid support for detection antibodies as well as a method to optimize the signal-to-noise ratio of fluorescence signals from E. coli. To improve the sensitivity of FACS measurements by eliminating artifactual signals from unrelated small particles in the samplemixture, E. coli with autodisplayed Z-domains was modified to express enhanced green fluorescence protein (eGFP) in the cytosol. By selectively gating the measurement channel for eGFP in the cytosol, we were able to discriminate between green fluorescent E. coli and artifact particles. We confirmed the efficacy of this optimization method for FACS-based immunoassay using several model reactions. Finally, as proof of concept, we used the FACS-based immunoassay and our optimization method for the medical diagnosis of human hepatitis B virus surface antigen.
Keyword Immunoassay, FACS, Z-domain, Medical diagnosis, Hepatitis B virus surface antigen
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