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• W1984782504 abstract "A cell-based, lawn format assay utilizing an in situ photocleavage method has been developed that allows the rapid examination of large bead-based compound libraries as discrete molecules. The format uses frog melanophore cells in a contiguous, adherent, confluent layer in small petri dishes covered with a 0.5–1-mm layer of agarose containing 130 micron diameter TentaGel beads at a density of 2–20 beads/mm2. Employing this technique a 9-mer, 442,368-member peptide library (designed around the 13 amino acid α-MSH peptide sequence) made up of 12 separate pools of 36,864 peptides/pool was assayed. Initially, a fraction (∼10%) of each pool was scanned (∼3700 beads from each pool) in 60-mm petri dishes to identify the most active pools. Upon direct photocleavage of the beads with UV light (365 nm), each petri dish was photographed over a 60-min period with a CCD camera to record changes in light intensity as an index of melanosome dispersion. Active beads were those that were surrounded by a localized decrease in light transmittance indicating melanosome dispersed cells. Upon examination with a dissecting microscope, single beads centrally located to a circular array of dispersed cells were identified and removed from the agarose and sequenced by Edman degradation to determine the peptide sequence. Re-synthesized peptides were re-examined against α-MSH receptor to confirm and quantify the activity. Several 9-mer peptides were identified with potencies similar to the natural 13-mer peptide. This method allows for the rapid screening of large bead-based photo-cleavable peptide libraries with the advantage that each compound is screened as a discrete molecule in a well-less format." @default.
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• W1984782504 date "1999-12-01" @default.
• W1984782504 modified "2023-09-26" @default.
• W1984782504 title "Use of a cell-based, lawn format assay to rapidly screen a 442,368 bead-based peptide library" @default.
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• W1984782504 doi "https://doi.org/10.1016/s1056-8719(00)00083-6" @default.
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