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PURIFICATION OF A FIBRINOLYSIS ENHANCING PROTEIN

Award Information

Agency:
Department of Health and Human Services
Branch:
N/A
Award ID:
19299
Program Year/Program:
1992 / SBIR
Agency Tracking Number:
19299
Solicitation Year:
N/A
Solicitation Topic Code:
N/A
Solicitation Number:
N/A
Small Business Information
ELCATECH, INC.
101 N CHESTNUT STREET Winston-salem, NC 27101
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Woman-Owned: No
Minority-Owned: No
HUBZone-Owned: No
 
Phase 1
Fiscal Year: 1992
Title: PURIFICATION OF A FIBRINOLYSIS ENHANCING PROTEIN
Agency: HHS
Contract: N/A
Award Amount: $50,000.00
 

Abstract:

DURING THE DEVELOPMENT OF OUR FIBRINOLYTIC ASSAY SYSTEM WE NOTED AN ENHANCEMENT OF FIBRINOLYTIC ACTIVITY IN PLASMA WHICH COULD NOT BE ACCOUNTED FOR BY ANY KNOWN FIBRINOLYTIC FACTOR. USING HEPARING AFFINITY CHROMATOGRAPHY, WITH A 0.5M NAC1 ELUTION AND AN IMMUNO AFFINITY COLUMN WE HAVE BEEN ABLE TO PARTIALLY PURIFY THIS FACTOR. THE IMMUNO AFFINITY COLUMN USES IMMOBILIZED MCAB-3, A MONOCLONAL ANTIBODY AGAINST A PLASMA ELUATE FROM A HEPARIN AFFINITY COLUMN. WE HAVE DEMONSTRATED THE ENHANCING ACTIVITY OF THE PROTEIN IN THE ENZYME LINKED FIBRINOLYTIC ASSAY (ELFA), A CLOT LYSIS ASSAY WITH ADDED T-PA, AND A BIO IMMUNOASSAY USING A TRANSFERABLE SOLID PHASE ENZYME LABELLED FIBRIN. USING THE BIO IMMUNOASSAY WE HAVE ESTIMATED PLASMA LEVELS TO BE APPROXIMATELY 4.5 UNITS. PLASMA PROTEIN LEVELS FOR THE COFACTOR APPEAR TO BE LESS THAN 50UG/ML. HPLC GEL PERMEATION CHROMATOGRAPHY REVEALS A MCAB-3 BINDING PROTEIN OF 78KD. THE PROTEIN COULD NOT BE ACTIVATED IN THE PRESENCE OF HIGH CONCENTRATIONS OF EXTRINSIC FIBRINOLYTIC ACTIVATORS, PRECLUDING THE POSSIBILITY OF PLASMINOGEN BEING RESPONSIBLE FOR THE ACTIVITY. THE PROTEIN DID NOT ACTIVATE PLASMINOGEN AND HAD NO MEASURABLE FIBRINOLYTIC ACTIVITY. IMMUNIDIFFUSION ANALYSIS OF THE INTACT PROTEIN REVEALED NON IDENTITY WITH VITRONECTIN OR HISTIDEINE RICH GLYCOPROTEIN, OTHER HEPARIN BINDING PLASMA PROTEINS IMPLICATED IN FIBRINOLYSIS MODULATION. WE WILL PURIFY THE PROTEIN TO HOMOGENEITY AND POSITIVELY IDENTIFY IT BY N-TERMINAL SEQUENCE ANALYSIS. WE WILL RAISE A POLYCLONAL ANTIBODY AGAINST THE PROTEIN TO USE IN THE DEVELOPMENT OF IMMUNOASSAYS FOR THE MEASUREMENT OF THE PROTEIN IN PLASMA.

Principal Investigator:

Mark X Triscott
9197773624

Business Contact:

Small Business Information at Submission:

Elcatech Inc
#64-1001 South Marshall Street Winston-salem, NC 27101

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