title: Flow cytometric immunoassay using E. coli with autodisplayed Z-domains.
abstract: Recently, we reported a highly sensitive immunoassay using Escherichia coli cells with autodisplayed Z-domains. In this work, E. coli cells with autodisplayed Z-domains were applied to the flow-cytometry-based 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. By using E. coli cells with only the autodisplayed Z-domains, immunoassay of multiple analytes could be performed simultaneously on the same sample. Flow cytometry can be used to identify the immunoassay type by simultaneously detecting the fluorescence signal from the cytosol (tdTomato) and the fluorescence from the outer membrane, enabling the quantification of bound analytes after treatment with additional fluorescently labeled antibodies. To demonstrate the immunoassay of multiple analytes by using flow cytometry, human hepatitis B virus surface antigen (HBsAg) and C-reactive protein (CRP), a broad spectrum inflammation marker, were used as model analytes.
authors.affiliation: Department of Materials Science and Engineering, Yonsei University, 50 Yonsei-Ro, Seodaemun-Gu, Seoul, 120-742, Republic of Korea.
authors.forename: Min
authors.initials: M
authors.lastname: Park
authors.forename: Ji-Hong
authors.initials: JH
authors.lastname: Bong
authors.forename: Gu
authors.initials: G
authors.lastname: Yoo
authors.forename: Joachim
authors.initials: J
authors.lastname: Jose
authors.forename: Min-Jung
authors.initials: MJ
authors.lastname: Kang
authors.forename: Jae-Chul
authors.initials: JC
authors.lastname: Pyun
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized; D002097:C-Reactive Protein / Q000032:analysis; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000276:immunology* / Q000378:metabolism*; D005434:Flow Cytometry / Q000379:methods*; D006041:Goats; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006801:Humans; D007118:Immunoassay / Q000379:methods*; D008164:Luminescent Proteins / Q000235:genetics
chemical_list: D055910:Antibodies, Immobilized; D006514:Hepatitis B Surface Antigens; D008164:Luminescent Proteins; D002097:C-Reactive Protein
keywords: Autodisplay; C-reactive protein; Flow cytometry; Hepatitis B virus surface antigen; Immunoassay; Z-domain