Yoo, Gu Bong, Ji-Hong Park, Min Kang, Min-Jung Jose, Joachim Pyun, Jae-Chul 2024-01-20T12:02:02Z 2024-01-20T12:02:02Z 2021-09-04 2013-07-10 0141-0229 https://pubs.kist.re.kr/handle/201004/127874 Escherichia coli cells with autodisplayed Z-domains could increase the sensitivity of immunoassays by immobilizing antibodies in a controlled orientation. In the work presented here, E. coli cells with autodisplayed Z-domains were immobilized to magnetic beads for subsequent immunoassay. In comparing conventional immunoassay using the E. coli cells with autodisplayed Z-domains, the magnetic-beadbased immunoassay improved immunoassay efficiency by minimizing the loss of E. coli cells during repeated centrifugation steps during washing. For the immobilization of E. coli cells to magnetic beads, the magnetic beads were modified with poly-L-lysine to bind to negatively charged E. coli cells. During the surface modification process, physical parameters such as the surface charge and size of the magnetic beads were analyzed to confirm the formation of E. coli-magnetic bead complexes. To test the feasibility of the magnetic-bead-based immunoassay, horseradish peroxidase (HRP) was used as a model analyte, and a biomarker for inflammatory diseases, C-reactive protein (CRP), was used for a demonstration of an application in medical diagnosis. (C) 2013 Elsevier Inc. All rights reserved. English ELSEVIER SCIENCE INC C-REACTIVE PROTEIN ESCHERICHIA-COLI ANTIBODY IMMOBILIZATION SPR BIOSENSOR RECOMBINANT PROTEINS SURFACE DISPLAY OUTER-MEMBRANE LAYER IMMUNOSENSOR ORIENTATION Magnetic-bead-based immunoassay using E. coli cells with autodisplayed Z-domains Article 10.1016/j.enzmictec.2013.03.022 1 ENZYME AND MICROBIAL TECHNOLOGY, v.53, no.2, pp.118 - 122 ENZYME AND MICROBIAL TECHNOLOGY 53 2 118 122 scie scopus 000321479700007 2-s2.0-84879017924 Biotechnology & Applied Microbiology Biotechnology & Applied Microbiology Article C-REACTIVE PROTEIN ESCHERICHIA-COLI ANTIBODY IMMOBILIZATION SPR BIOSENSOR RECOMBINANT PROTEINS SURFACE DISPLAY OUTER-MEMBRANE LAYER IMMUNOSENSOR ORIENTATION Autodisplay Z-domain Magnetic bead Immunoassay Surface modification