Molecular pincers: antibody-based homogeneous protein sensors.
We describe here a new homogeneous antibody-based protein sensor design (molecular pincers) that allows rapid and sensitive detection of a specific protein in solution. In the presence of the target protein these sensors produce fluorescence signal derived from target-dependent annealing of short complementary fluorochrome-labeled oligonucleotides attached to a pair of target-specific antibodies via nanometer-scale flexible linkers. The sensors allow near-instantaneous detection of the target with sensitivity and specificity approaching that of enzyme-linked immunosorbent assay (ELISA) but requiring no sample manipulation other then the addition of the sample to the sensor mix. We used cardiac troponin I and C-reactive protein as the targets to validate these desirable properties of the sensors. Due to the availability of antibodies to thousands of interesting targets and the straightforward design blueprint of the sensors we expect their wide-ranging applications in research and medical diagnosis, especially when simplicity, high throughput, and short detection time are essential.
Heyduk
Ewa
E
Edward A. Doisy Department of Biochemistry and Molecular Biology, St. Louis University Medical School, 1100 South Grand Boulevard, St. Louis, Missouri 63104, USA. heyduk@slu.edu
Dummit
Benjamin
B
Chang
Yie-Hwa
YH
Heyduk
Tomasz
T
D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D000918:Antibody Specificity; D015374:Biosensing Techniques / Q000379:methods*; D002097:C-Reactive Protein / Q000032:analysis / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D031541:Fluorescence Resonance Energy Transfer / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D016328:NF-kappa B / Q000032:analysis / Q000276:immunology; D009841:Oligonucleotides / Q000737:chemistry; D011506:Proteins / Q000032:analysis* / Q000276:immunology; D019210:Troponin I / Q000032:analysis / Q000276:immunology; D016159:Tumor Suppressor Protein p53 / Q000032:analysis / Q000276:immunology
D000911:Antibodies, Monoclonal; D005456:Fluorescent Dyes; D016328:NF-kappa B; D009841:Oligonucleotides; D011506:Proteins; D019210:Troponin I; D016159:Tumor Suppressor Protein p53; D002097:C-Reactive Protein