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 antibody. The sensitivity and specificity is enhanced 28-fold by incorporation of aFPs into solid-phase surface.
Zhong
Joan Q
JQ
Whitehead Institute for Biomedical Research, MIT, Nine Cambridge Center, Cambridge, MA 02142, USA.
Freyzon
Yelena
Y
Ehrlich
Daniel J
DJ
Matsudaira
Paul
P
D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D015374:Biosensing Techniques; D005453:Fluorescence; D049452:Green Fluorescent Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006388:Hemagglutinins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011485:Protein Binding / Q000235:genetics; D015202:Protein Engineering; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence; D013499:Surface Properties
D000911:Antibodies, Monoclonal; D006388:Hemagglutinins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins