Current methods for the detection of diagnostic proteins entail time- and resource-intensive immunochemical techniques that, even under ideal circumstances, require an hour or more to return an answer to the clinician's hands. This sluggish response fits poorly into the timeframe of modern healthcare, hindering patient compliance and, in some circumstances, the efficiency and safety with which drugs and procedures are administered. (Consider: a diagnostic test that returns an answer in less than 15 minutes differs qualitatively from a test that requires 30 minutes because the latter requires a second interaction with the clinician.) Here we propose the development of a rapid, point-of-care method for the simultaneous detection of multiple diagnostic proteins. Our approach, which utilizes electrochemistry to monitor the binding-induced folding of polypeptide- or protein-based recognition elements, will be rapid, specific, convenient and, critically, selective enough to employ directly in blood serum and -we propose- whole blood. And while our initial development efforts will focus on the detection of proteins diagnostic of HIV infection (such as anti-HIV antibodies), the approach will be general enough to be of use in the detection of a wide range of clinically relevant markers. The development of rapid, parallelizable point-of-care diagnostics could significantly impact the safety, compliance and efficacy of therapies and medical procedures ranging from the detection of infectious diseases, the treatment of autoimmune diseases, and the routine monitoring of health status.

Public Health Relevance

Here we proposed the development of a reagentless, electrochemical platform for the simultaneous detection of multiple proteins diagnostic of disease. The proposed technology will be rapid, specific, and selective enough to employ directly at the point of care, thus significantly improving the speed with which molecular diagnostics can be performed and their results acted upon. This, in turn, will improve the safety, compliance and efficacy of therapies and procedures ranging from the detection of infectious diseases, the treatment of autoimmune diseases, and the routine monitoring of health status.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
5R01EB002046-07
Application #
7591704
Study Section
Enabling Bioanalytical and Biophysical Technologies Study Section (EBT)
Program Officer
Korte, Brenda
Project Start
2001-05-01
Project End
2011-12-31
Budget Start
2009-04-01
Budget End
2010-03-31
Support Year
7
Fiscal Year
2009
Total Cost
$238,613
Indirect Cost
Name
University of California Santa Barbara
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
094878394
City
Santa Barbara
State
CA
Country
United States
Zip Code
93106
Dingledine, Raymond; Coulter, Douglas A; Fritsch, Brita et al. (2017) Transcriptional profile of hippocampal dentate granule cells in four rat epilepsy models. Sci Data 4:170061
White, Ryan J; Kallewaard, Hannah M; Hsieh, Wen et al. (2012) Wash-free, electrochemical platform for the quantitative, multiplexed detection of specific antibodies. Anal Chem 84:1098-103
Yoo, Tae Yeon; Meisburger, Steve P; Hinshaw, James et al. (2012) Small-angle X-ray scattering and single-molecule FRET spectroscopy produce highly divergent views of the low-denaturant unfolded state. J Mol Biol 418:226-36
Rowe, Aaron A; Bonham, Andrew J; White, Ryan J et al. (2011) CheapStat: an open-source, ""do-it-yourself"" potentiostat for analytical and educational applications. PLoS One 6:e23783
Rowe, Aaron A; White, Ryan J; Bonham, Andrew J et al. (2011) Fabrication of electrochemical-DNA biosensors for the reagentless detection of nucleic acids, proteins and small molecules. J Vis Exp :
Rowe, Aaron A; Chuh, Kelly N; Lubin, Arica A et al. (2011) Electrochemical biosensors employing an internal electrode attachment site and achieving reversible, high gain detection of specific nucleic acid sequences. Anal Chem 83:9462-6
Plaxco, Kevin W; Soh, H Tom (2011) Switch-based biosensors: a new approach towards real-time, in vivo molecular detection. Trends Biotechnol 29:1-5
Lubin, Arica A; Plaxco, Kevin W (2010) Folding-based electrochemical biosensors: the case for responsive nucleic acid architectures. Acc Chem Res 43:496-505
Lawrence, Camille; Kuge, Jennifer; Ahmad, Kareem et al. (2010) Investigation of an anomalously accelerating substitution in the folding of a prototypical two-state protein. J Mol Biol 403:446-58
Vallee-Belisle, Alexis; Plaxco, Kevin W (2010) Structure-switching biosensors: inspired by Nature. Curr Opin Struct Biol 20:518-26

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