The use of biotin labeled ligands in receptor binding assays is proposed as a substitute for radiolabled ligands. High affinity biotinylated ligands are used in immunochemistry studies but are not generally employed in receptor binding experiments. Biotinylated ligands also have high affinities for the proteins avidin and streptavidin. These proteins can be coupled with various enzymes which catalyze spectrophotometric, fluorometric, or chemiluminescent detection in microliter plates. Using standard receptor binding protocols, biotinylated ligands can be incubated with the appropriate receptor and possible competitors for the receptor. Free or receptor bound biotinylated ligands are detected by binding to the avidin or st reptavidin enzyme conjugates. This mechanism makes it possible for one molecule of biotinylated ligand to be responsible for the generation of many molecules of product, providing an amplification of the initial signal. Several strategies for selected receptor binding assays, utilizing biotinylated ligands and a microliter plate assay format assays will be examined. The knowledge gained in these investigations should aid in the development of additional high throughput nonradioactive receptor binding assays.

Agency
National Institute of Health (NIH)
Institute
National Institute of Mental Health (NIMH)
Type
Small Business Innovation Research Grants (SBIR) - Phase I (R43)
Project #
1R43MH053461-01
Application #
2253581
Study Section
Small Business Research Review Committee (MHSB)
Project Start
1995-05-01
Project End
1995-10-31
Budget Start
1995-05-01
Budget End
1995-10-31
Support Year
1
Fiscal Year
1995
Total Cost
Indirect Cost
Name
Analytical Biological Services, Inc.
Department
Type
DUNS #
612501502
City
Wilmington
State
DE
Country
United States
Zip Code
19801