E. coli outer membrane with autodisplayed Z-domain as a molecular recognition layer of SPR biosensor.
In this work, the Z-domain of protein A was expressed as a recombinant-fusion protein on the outer membrane of Escherichia coli by using "Autodisplay" technology, and the outer membrane with Z-domains was layered on a SPR biosensor as a highly sensitive molecular recognition layer. The outer membrane layer formation was demonstrated by using various surfaces with controlled hydrophobicity. The effect of orientation control by the outer membrane with Z-domains was estimated to have far higher sensitivity and far improved limit of detection (LOD) in comparison to the conventional immunoassay by using a model analyte, horseradish peroxidase (HRP). The SPR biosensor with the outer membrane layer was applied to the detection of C-reactive protein (CRP). From these experiments, the LOD of SPR biosensor was estimated to improve more than 100-fold compared to the SPR biosensor with the antibody-layer by physical adsorption.
Jose
Joachim
J
Institute of Pharmaceutical Chemistry, Heinrich Heine University, Duesseldorf, Germany.
Park
Min
M
Pyun
Jae-Chul
JC
D001681:Biological Assay / Q000295:instrumentation*; D015374:Biosensing Techniques / Q000295:instrumentation; D002097:C-Reactive Protein / Q000032:analysis*; D002462:Cell Membrane / Q000378:metabolism*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D004926:Escherichia coli / Q000378:metabolism*; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism*; D020349:Surface Plasmon Resonance / Q000295:instrumentation*
D013205:Staphylococcal Protein A; D002097:C-Reactive Protein