Enhanced detection sensitivity using a novel solid-phase incorporated affinity fluorescent protein biosensor

We engineered green fluorescent protein (GFP) into affinity fluorescent proteins (aFPs) biosensors. The aFPs detect protein–protein interactions by enhanced fluorescence intensity. In a proof of principle demonstration, aFPs containing haemagglutinin (HA) tag bind specifically to the anti-HA antibod...

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Veröffentlicht in:	Biomolecular engineering 2004-04, Vol.21 (2), p.67-72
Hauptverfasser:	Zhong, Joan Q., Freyzon, Yelena, Ehrlich, Daniel J., Matsudaira, Paul
Format:	Artikel
Sprache:	eng
Schlagworte:	Affinity fluorescent protein Antibodies, Monoclonal - chemistry Antibodies, Monoclonal - metabolism Binding Sites - genetics Biosensing Techniques Biosensor Enhanced fluorescence intensity Fluorescence Green fluorescent protein Green Fluorescent Proteins - chemistry Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism Hemagglutinins - chemistry Hemagglutinins - genetics Hemagglutinins - metabolism Protein Binding - genetics Protein Engineering Protein Structure, Tertiary Recombinant Fusion Proteins - chemistry Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Sensitivity and Specificity Solid-phase assay Spectrometry, Fluorescence Surface Properties
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Beschreibung
Zusammenfassung:	We engineered green fluorescent protein (GFP) into affinity fluorescent proteins (aFPs) biosensors. The aFPs detect protein–protein interactions by enhanced fluorescence intensity. In a proof of principle demonstration, aFPs containing haemagglutinin (HA) tag bind specifically to the anti-HA antibody. The sensitivity and specificity is enhanced 28-fold by incorporation of aFPs into solid-phase surface.
ISSN:	1389-0344 1878-559X
DOI:	10.1016/j.bioeng.2003.10.004