Enzyme Microb Technol
September 2019
α2,3-Sialyltransferase from Pasteurella multocida (PmST1) is an enzyme that transfers a sialyl group of donor substrates to an acceptor substrate called N-acetyl-d-lactosamine (LacNAc). In this study PmST1 was expressed on the outer membrane of wildtype Escherichia coli (BL21) with lipopolysaccharide (LPS) and ClearColi with no LPS, and then the enzyme activity and expression level of PmST1 were compared. As the first step, the expression levels of PmST1 on the outer membranes of wildtype E.
View Article and Find Full Text PDFThe autoimmune diseases systemic lupus erythematosus (SLE) and Sjogren's syndrome (SS) are diagnosed by detection of autoantibodies against Ro and La protein autoantigens, respectively. In this work, the diagnosis of autoimmune diseases SLE and SS was demonstrated using thermophoresis of Escherichia coli with the autodisplayed autoantigens. Ro and La protein autoantigens were autodisplayed by constitutive expression together with a fluorescent protein called tdTomato in the cytosol.
View Article and Find Full Text PDFThe objective of this study was to present an immunoassay for the diagnosis of Sjögren's syndrome based on the autodisplayed La/SSB protein on the outer membrane of intact E. coli (strain UT-5600) and LPS-free E. coli (ClearColi™).
View Article and Find Full Text PDFThe autodisplay technology has been applied for expression of a desired protein on the outer membrane (OM) of Escherichia coli. In this work, the OM fractions of E. coli with two autodisplayed proteins were separately prepared and mixed to demonstrate the feasibility of control over the ratio of two autodisplayed proteins.
View Article and Find Full Text PDFEscherichia coli cells with autodisplayed Z-domains have been used for immunoassays of specific target analytes. In this study, a magnetite suspension was used for the washing step in immunoassays of E. coli cells with autodisplayed Z-domains.
View Article and Find Full Text PDFBiochim Biophys Acta
December 2015
In this work, two proteins, Z-domains and bovine casein, were auto-displayed on the outer membrane of the same Escherichia coli cells by co-transformation of two different auto-display vectors. On the basis of SDS-PAGE densitometry, Z-domains and bovine casein were expressed at 3.12 × 10⁵ and 1.
View Article and Find Full Text PDFIn this work, adrenodoxin (Adx) was expressed on the outer membrane of E. coli by autodisplay and then the iron-sulfur cluster was incorporated into apo-Adx by an anaerobic reconstitution process. For the determination of the redox potentials of the iron-sulfur clusters of the autodisplayed Adx, E.
View Article and Find Full Text PDF"Autodisplay technology" is an expression technique used to display the various recombinant proteins on the outer membrane (OM) of Escherichia coli. The resulting autodisplayed Z-domain has been used to improve the sensitivity of immunoassays. In this work, a facile isolation method of the OM fraction of E.
View Article and Find Full Text PDFEnzyme Microb Technol
November 2014
The outer membrane of Escherichia coli was previously isolated as a liposome-like outer membrane particle using an enzymatic treatment for lysozymes; for immunoassays, the particles were subsequently layered on solid supports via hydrophobic interactions. This work presents an enzyme-free isolation method for the E. coli outer membrane with autodisplayed Z-domains using ultrasonication.
View Article and Find Full Text PDFA microarray-based immunoassay for the detection of autoantibodies against Ro protein was developed using Escherichia coli with autodisplayed Ro proteins (Ro(+)-E. coli). Patient serum usually contains various antibodies against the outer membrane components of E.
View Article and Find Full Text PDFWe have developed a carbon nanotube (CNT) film-based biosensor with a metal semiconductor field effect transistor structure (MESFET). A gold top gate was deposited on the middle of the CNT channel and probe antibodies were immobilized on the gold top gate with an antibody-binding protein, protein G or Escherichia coli outer membrane (OM) with autodisplayed Z-domains of protein A. These CNT-MESFET biosensors exhibited a higher sensitivity than the CNT-FET biosensor with probe antibodies immobilized using a chemical linker, since the orientation of immobilized antibodies was controlled by the antibody-binding proteins.
View Article and Find Full Text PDFEnzyme Microb Technol
August 2013
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.
View Article and Find Full Text PDFEnzyme Microb Technol
July 2013
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.
View Article and Find Full Text PDFEscherichia coli with autodisplayed Z-domains was reported to improve the sensitivity of immunoassays by the orientation control of antibodies. In this work, a sensitive microplate-based immunoassay is presented by immobilizing E. coli cells to a surface-modified microplate.
View Article and Find Full Text PDF