Optimization of a FACS based-immunoassay using E. coli autodisplaying Z-domains

被引:0
|
作者
Min Park
Ji-Hong Bong
Gu Yoo
Joachim Jose
Min-Jung Kang
Jae-Chul Pyun
机构
[1] Yonsei University,Department of Materials Science and Engineering
[2] University of Muenster,Institute of Pharmaceutical and Medical Chemistry
[3] Korea Institute of Science and Technology (KIST),undefined
来源
BioChip Journal | 2013年 / 7卷
关键词
Immunoassay; FACS; Z-domain; Medical diagnosis; Hepatitis B virus surface antigen;
D O I
暂无
中图分类号
学科分类号
摘要
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 sample mixture, 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.
引用
收藏
页码:173 / 179
页数:6
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