Rapid homogeneous antibody-based detection of proteins

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$129,493.00
Award Year:
2007
Program:
STTR
Phase:
Phase I
Contract:
1R41GM079891-01
Award Id:
85692
Agency Tracking Number:
GM079891
Solicitation Year:
n/a
Solicitation Topic Code:
n/a
Solicitation Number:
n/a
Small Business Information
815 WENNEKER DRIVE, SAINT LOUIS, MO, 63124
Hubzone Owned:
N
Minority Owned:
N
Woman Owned:
N
Duns:
100512883
Principal Investigator:
TOMASZHEYDUK
(314) 977-9238
HEYDUKT@SLU.EDU
Business Contact:
MARGARETMENG
() -
yiechang@mediomics.com
Research Institute:
SAINT LOUIS UNIVERSITY

221 N GRAND BLVD
SAINT LOUIS, MO, 63103 2194

Nonprofit college or university
Abstract
DESCRIPTION (provided by applicant): Detection of the presence and the determination of the amount of a specific protein using various techniques utilizing antibodies specific to the target protein are one of the most important tools in biomedical research , medical diagnosis and detection of biological threat agents. These techniques (such as, for example, ELISA) take advantage of the exquisite specificity of antigen binding by the antibodies and constitute the current gold-standard of specific protein dete ction. While extremely useful, these existing techniques have some limitations. One important limitation is relatively long time required to perform the assay. This limitation is especially important in situations where the answers need to be known immedia tely such as, for example, in determination of biomarkers important for emergency room care decisions. Another important limitation is a relative technical complexity of these assays which limits their point-of-care applicability and their adaptation and t ailoring to the needs of the intended end-users and their settings. The major goal of this project will be to develop a new antibody-based protein detection methodology (molecular pincers), which will retain all advantages of classical antibody-based assay s while reducing the time, complexity, and the cost of the assay. This new methodology is an extension of nucleic-acid-based molecular beacon methodologies previously developed by us. This new assay format is homogeneous requiring no complicated sample m anipulations. A sample needs only to be added to the assay mixture and after a short incubation an easy to read optical signal (fluorescence) reports the presence of the target molecule. While the molecular pincers will be applicable to any situation invol ving antibody-based detection of antigens, their unique properties and characteristics make them especially applicable in situations requiring rapid results. In Phase I of this project we will use cardiac troponin, an excellent example of a cardiac biomark er requiring quick determination in emergency room in patients with chest pains, as a model target protein to test the concept of molecular pincers. While all of the proposed research in Phase I will involve cardiac troponin as a model, the lessons related to molecular pincer design will be generally applicable to other systems. Following successful completion of Phase I, in Phase II molecular pincers to an extensive panel of proteins will be developed, tested and validated. A priority in Phase II will be g iven to target proteins with an established need for their rapid determination. Additional goal of Phase II will be to develop signal amplification procedures compatible with molecular pincer design to extend sensitivity of the assay. In Phase I of this pr oposal we will test and validate two designs of molecular pincers: Aim #1: To develop and test molecular pincer assay for detecting troponin based on two antibodies recognizing two nonoverlaping epitopes of troponin. Aim #2: To develop and test single anti body-based molecular pincer assay for detecting troponin based on competition between the protein and the epitope-containing synthetic peptide. Successful completion of the above aims will validate the practical feasibility of molecular pincer design. We e xpect that molecular pincers will find very broad applications as a diagnostic tool allowing rapid and straightforward tests for biomarkers of human disease and as a research tool replacing, when possible, ELISA tests with a much simpler to perform homogen eous assay.

* information listed above is at the time of submission.

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