Z-domains Autodisplaying E. coli for Flow cytometry-based Immunoassays
- Z-domains Autodisplaying E. coli for Flow cytometry-based Immunoassays
- 박민; 변재철; Joachim Jose; 강민정
- autodisplay; Z-domain; Flow cytometry; immunoassay
- Issue Date
- 2014 한국바이오칩학회 춘계학술대회
- Fluorescence-activated cell sorter (FACS)-based immunoassays using E. coli cells with autodisplayed Z-domains were performed to (1) improve the sensitivity of the immunoassay through orientation control of antibodies with autodisplayed Z-domains and (2) detect multiple analytes with one measurement. IgG-binding Z-domains were expressed on the surface of E. coli by autodisplay technology. As FACS measures the fluorescent signal from individual E. coli cell, the optimal parameters were determined, and the minimum amount of analyte required for the E. coli cell-based immunoassay was identified. The diagnosis method for acute myocardinal infarction with troponin-I was developed. E. coli cells with autodisplayed Z-domains were applied to the simultaneous detection of multiple analytes. The E. coli cells were doubly transfected to express a fluorescent protein (tdTomato) in the cytosol and the autodisplayed Z-domains on the outer membrane. Flow cytometry can be used to identify analytes by simultaneously detecting the fluorescence signal from the cytosol. The fluorescence from the outer membrane enabled the quantification of bound analytes after treatment with additional fluorescently labeled antibodies.
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