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Ultrasensitive pathway phosphoproteomics

Award Information

Agency:
Department of Health and Human Services
Branch:
N/A
Award ID:
89158
Program Year/Program:
2008 / SBIR
Agency Tracking Number:
GM081956
Solicitation Year:
N/A
Solicitation Topic Code:
N/A
Solicitation Number:
N/A
Small Business Information
AMPLIFIED PROTEOMICS, INC.
AMPLIFIED PROTEOMICS, INC. 2462 WYANDOTTE ST MOUNTAIN VIEW, CA 94043 2313
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Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No
 
Phase 1
Fiscal Year: 2008
Title: Ultrasensitive pathway phosphoproteomics
Agency: HHS
Contract: 1R43GM081956-01A1
Award Amount: $194,424.00
 

Abstract:

DESCRIPTION (provided by applicant): Site-specific protein phosphorylation is essential for signal transduction in mammalian cells, and perturbation in phosphoprotein levels is a hallmark of cancerous transformation. Unfortunately there are no sensitive me thods for detecting multiple phosphorylation events in parallel, so researchers are forced to run multiple Western blot or ELISA experiments, expending precious samples, in order to dissect these signaling pathways. We describe here a novel, high throughpu t, ultrasensitive method for detecting multiple phosphoproteins simultaneously. The method consists of a sandwich immuno- assay comprised of multiple pan-specific capture antibodies and a cocktail of detection antibodies against both phosphorylated and non -phosphorylated epitopes of the target proteins. Each detection antibody is labeled with a unique DNA tag. Real-time PCR is then used to quantify all of the DNA tags in parallel, thus revealing the quantities of the cognate (phospho-)proteins in the biolog ical samples. We show that this immuno-PCR method is more sensitive than ELISA and can detect endogenous proteins at sub-picomolar levels in real biological samples. In this grant application, we propose to develop and validate this concept by developing a highly sensitive, multiplexed assay to measure the phosphorylation states of signaling proteins in the BCR-ABL pathway. This platform will allow researchers to measure numerous phosphoproteins from minimal (~50 ul) sample volumes. Because all steps use mi crotiter plates and simple manipulations, this technology will allow researchers, with minimal effort and standard equipment, to decipher changes in phosphorylation patterns resulting from numerous biological stimuli.

Principal Investigator:

Business Contact:


ciwang_99@yahoo.com
Small Business Information at Submission:

AMPLIFIED PROTEOMICS, INC.
AMPLIFIED PROTEOMICS, INC. 2462 WYANDOTTE ST MOUNTAIN VIEW, CA 94043

EIN/Tax ID: 120406850
DUNS: N/A
Number of Employees: N/A
Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No