SERUM ERYTHROPOIETIN BIOASSAY USING A NEW CELL LINE

Award Information
Agency: Department of Health and Human Services
Branch: N/A
Contract: N/A
Agency Tracking Number: 16739
Amount: $499,719.00
Phase: Phase II
Program: SBIR
Awards Year: 1993
Solicitation Year: N/A
Solicitation Topic Code: N/A
Solicitation Number: N/A
Small Business Information
Merrifield Research Laboratory
2723-b Merrilee Drive, Fairfax, VA, 22031
DUNS: N/A
HUBZone Owned: N
Woman Owned: N
Socially and Economically Disadvantaged: N
Principal Investigator
 Hankins, David W
 Principal Investigator
 (703) 560-1910
Business Contact
 1 R43 HL47259-01
Phone: () -
Research Institution
N/A
Abstract
WE PLAN TO EXPLOIT THE ISOLATION OF A CELL LINE FROM ERYTHROLEUKEMIC MURINE TUMORS WHICH IS HIGHLY SENSITIVE LINE FROM ERYTHROLEUKEMIC MURINE TUMORS WHICH IS HIGHLY SENSITIVE TO ERYTHROPOIETIN. WE WILL UTILIZE THIS LINE TO DEVELOP A SENSITIVE AND SPECIFIC BIOASSAY FOR ERYTHROPOIETIN IN HUMAN SERUM. OUR PRELIMINARY WORK HAS SHOWN THAT WE HAVE ISOLATED A LINE WHICH IS SENSITIVE TO VERY LOW LEVELS OF ERYTHROPOIETIN. THIS LINE, HC24 HAS NOW BEEN CARRIED CONTINUOUSLY FOR MORE THAN 120 PASSAGES OVER AN 18 MONTH PERIOD. IT DOES NOT APPEAR TO BE SENSITIVE TO A VARIETY OF OTHER GROWTH FACTORS INCLUDING IL-1, IL-2, IL-3, GM-CSF OR BY WORD OF MOUTH (DR. S. RUSCETTI) IL-4, IL-5, IL-6 AND G-CSF. THE DESIGN OF THIS SPECIFIC STUDY IS TO ISOLATE AND CHARACTERIZE STABLE, HIGHLY EPO SENSITIVE CLONAL CELL LINES FROM THE PARENTERAL HC LINES. WE WILL USE A TRITIATED THYMIDINE UPTAKE PROCEDURE DESCRIBED BY KRYSTAL TO ASSAY RESPONSE TO ERYTHROPOIETIN BUT WILL ALSO EVALUATE THE POSSIBILITY OF USING AN MTT SUBSTRATE ASSAY TO REPLACE TRITIATED THYMIDINE. WE WILL COMPARE THESE METHODS AND WILL DETERMINE THE FEASIBILITY OF ASSAYING BIOLOGIC ACTION OF EPO IN-VITRO IN THE PRESENCE OF HUMAN SERUM. WE WILL LOOK FOR THE EFFECTS OF HUMAN SERUM ON INHIBITING CELLS AND WILL USE ANTIBODY DEPLETION OF SAMPLES REMOVING EPO TO SEE WHETHER OR NOT ACTIVITY IS REMOVED. WE WILL LOOK FOR THE EFFECTS OF HUMAN SERUM ON QUANTITATING EPO AND ANALYZE THE ABILITY TO DETECT VERY LOW LEVELS IN VERY SMALL VOLUMES.

* information listed above is at the time of submission.

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