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12th International Symposium on Bioluminescence & Chemiluminescence |
Symposium abstracts:
Van Arsdell, Scott W., Murphy, Kevin P., Orencole, Scott F., Pande, Rajiv, Vietz, Christine M., Burns , Christine
Pierce Boston Technology Center, Inc, 30 Commerce Way, Woburn, MA 01801-1059, USA
We have developed a "mini-array" system suitable for the high-throughput quantification of proteins. The mini-array is produced by spotting a 3 x 3 pattern of 9 different monoclonal antibodies (mAbs) in the bottoms of the wells of 96-well polystyrene plates. Each 96-well plate can produce data from 864 array features (9 x 96). Target proteins are captured by the arrayed mAbs, and then detected in a sandwich ELISA format using biotinylated mAbs followed by the addition of a streptavidin-horseradish peroxidase (HRP) conjugate and a chemiluminescent substrate. The light produced from the HRP catalyzed oxidation of the substrate at each spot in the array is measured by imaging the entire plate with a commercially available CCD camera. A cocktail of the target proteins is run on the plate as a standard curve (typically 0.8 to 400 pg/ml). The concentrations of the proteins in the samples are calculated by comparing the intensity of the spots in the wells containing the samples to the intensity of the spots in the wells containing the standard curve. Array panels have been developed for the quantification of human inflammatory cytokines, Th1/Th2 cytokines, and chemokines. Panels have also been developed for quantification of mouse cytokines.
This
is a preprint of an article accepted for publication in Luminescence: Copyright
2001 John
Wiley & Sons, Ltd (Wiley website)