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A novel drug detection assay using fluorescent biosensor technology

Award Number: R41GM110813
ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES
OPDIV: NIH
AWARD CLASS: DISCRETIONARY
AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)
PERIOD OF PERFORMANCE START DATE: 01/15/2015
PERIOD OF PERFORMANCE END DATE: 01/14/2017

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A novel drug detection assay using fluorescent biosensor technology - ¿ DESCRIPTION (provided by applicant): This phase 1 project has as its overall objectives: to design, engineer, optimize and implement a novel, genetically encoded, fluorescent drug sensor (FDS) based on Förster Resonance Energy Transfer (FRET), by exploiting conformational rearrangements induced by drug interactions with human serum albumin (SA). SA is uniquely suited for a general drug sensor because it possesses multiple binding sites for small molecules. We have acquired the ORF (open reading frame) encoding human SA, and will insert this ORF into Gateway vectors, purify the chimeric proteins from E. coli and analyze fluorescence responses to drugs using a 96-well microplate spectrofluorimeter. During the first year we will focus on design and construction of sensors; once a successful sensor has been obtained, we will optimize and fine-tune the sensor to different ligand groups. The following specific aims are proposed to achieve the goals of phase I of this project: 1. Convert the SA gene into Gateway-compatible format, insert into a suite of Gateway vectors (~60), transform E. coli and grow cultures, test responses in crude lysates to drugs, and monitor spectra of promising affinity-purified sensor proteins. 2. Obtain binding isotherms and determine affinities, compare to published data, for drug binding to SA constructs in comparison with standard SA-fluorescein. 3. Optimize signal to noise ratio by linker mutagenesis and create a series of specificity and affinity mutants. 4. Screen NCI Oncology Drug Set (124 compounds) to confirm sensor's general sensing capacity. Development of a successful new drug requires identifying those that show maximal ability to reach target cells (permeability) while having minimal effects on drug transporters (drug:drug interactions). For this reason, the Fluorosome division of GLSynthesis Inc. has established collaboration with Prof. W. Frommer at the Carnegie Institution for Science, Stanford University, to create genetically encoded FRET drug sensors (FDS) from SA. This novel drug sensor technology is expected to enhance the commercial use of Fluorosome(r)-based assays and lead to the development of novel assay systems.


Issue Date FY	Funding FY	Legal Entity Name	Legal Entity Address	Legal Entity City	Legal Entity State	Legal Entity Zip Code	Legal Entity COUNTY	Legal Entity COUNTRY	Assistance Listing	Award Code	Budget Year	Action Date	Action Type	Action Amount

Issue Date FY: 2017 ( Subtotal = $0 )
2017	2015	GLSYNTHESIS INC.	298 HIGHLAND ST	WORCESTER	MA	01602	WORCESTER	USA	Biomedical Research and Research Training	000	1	4/12/2017	NEW	$0
														Subtotal = $0

Issue Date FY: 2015 ( Subtotal = $320,466 )
2015	2015	GLSYNTHESIS, INC	ONE INNOVATION DRIVE	WORCESTER	MA	01605	WORCESTER	USA	Biomedical Research and Research Training	000	1	1/9/2015	NEW	$320,466
														Subtotal = $320,466

Grand Total All Awards = $320,466

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A novel drug detection assay using fluorescent biosensor technology

Award Number: R41GM110813

ORGANIZATION: NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES

OPDIV: NIH

AWARD CLASS: DISCRETIONARY

AWARD ACTIVITY TYPE: SCIENTIFIC/HEALTH RESEARCH (INCLUDES SURVEYS)

PERIOD OF PERFORMANCE START DATE: 01/15/2015

PERIOD OF PERFORMANCE END DATE: 01/14/2017

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