Escherichia coli with autodisplayed Z-domain of protein A for signal amplification of SPR biosensor
- Escherichia coli with autodisplayed Z-domain of protein A for signal amplification of SPR biosensor
- Joachim Jose; 정지원; Byoung-Jin Jeon; Ruth Maria Maas; 남창훈; 변재철
- Autodisplay; SPR biosensor; Myoglobin; Immunosensor; Signal amplification
- Issue Date
- Biosensors & bioelectronics
- VOL 24, NO 5, 1324-1329
- Generally, an immunoaffinity SPR biosensor detects a target analyte in a sample through highly selective adsorption by using the antigen–antibody interaction. For improving the sensitivity, various kinds of particles have been added to the already bound analytes on the SPR biosensor (sandwich assay). In this work, signal amplification was demonstrated by the expression of the IgG-binding Z-domain of protein A on the outer membrane of Escherichia coli via “Autodisplay”. The amount of Z-domain of protein A expressed on the outer membrane was calculated to be 280,000 molecules per cell. In addition, the IgGbinding ability of the expressed proteinwas characterized using FACS analysis. The signal amplification of the SPR biosensor was performed in the sandwich assay format using a model of horseradish peroxidase (HRP); the limit of detectionwas determined to be significantly improved from1 g/ml to 1 ng/ml. Finally, myoglobin analysis was demonstrated for the medical diagnosis of cardiac diseases. The detection limit was estimated to be improved from 10 ng/ml to <1 ng/ml. These results showthat Z-domain-displaying E. coli can be successfully used for the signal amplification of immunoaffinity biosensors, thereby improving the sensitivity and the limit of detection.
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