Cytokines serve a variety of regulatory functions, one of the most important being the regulation of T-cell differentiation. Secreted cytokine profiles have been demonstrated to be predictive of different T-cell differentiation, especially cytokines regulating cell-mediated and humoral immunity. In order to measure different cytokine profiles simultaneously in individual samples, a basic cytokine chip has been developed based on solid-phase immunoaffinity extraction of the analytes of interest. Arrays of recombinant streptavidin spots were derivatized to the surfaces of silanized glass microscope slides via a carbonyldiimidazole bridge. Anti-cytokine antibodies were biotinylated via their carbohydrate moieties and bound the the steptavidin. Samples were labeled with a laser dye prior to analysis and incubated with the chip. Following removal of non-reactive materials by washing, the bound analytes were read in a scanning laser densitometer. Sensitivity of the chip was found to be 100 - 150 pg/ml for all five cytokines studied. Further studies require optimization of the chip sensor and reading mechanism to further increase sensitivity. Selectivity studies indicated that the chip was capable of a high degree of specificity when using mixed cytokine samples. Development of new, increased selectivity chips open the door to studying a number of important regulatory pathways such as hematopoesis, inflammation neurogenic regulation, and wound healing simulataneously on the same chip. Future models are expected to contain 30-100 ligands making multiple analyses possible.

Agency
National Institute of Health (NIH)
Institute
Office of The Director, National Institutes of Health (OD)
Type
Intramural Research (Z01)
Project #
1Z01OD011029-01
Application #
6413434
Study Section
(DBEP)
Project Start
Project End
Budget Start
Budget End
Support Year
1
Fiscal Year
2000
Total Cost
Indirect Cost
Name
Office of the Director, NIH
Department
Type
DUNS #
City
State
Country
United States
Zip Code