期刊
METHODS AND APPLICATIONS IN FLUORESCENCE
卷 1, 期 1, 页码 -出版社
IOP PUBLISHING LTD
DOI: 10.1088/2050-6120/1/1/017001
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资金
- Medical Research Council [MC_G0802522] Funding Source: Medline
- MRC [MC_G0802522] Funding Source: UKRI
- Medical Research Council [MC_G0802522] Funding Source: researchfish
One-bead one-compound combinatorial library beads exhibit varying levels of autofluorescence after solid phase combinatorial synthesis. Very often this causes significant problems for automated on-bead screening using TentaGel beads and fluorescently labeled target proteins. Herein, we present a method to overcome this limitation when fluorescence activated bead sorting is used as the screening method. We have equipped the COPAS bead sorting instrument with a high-speed profiling unit and developed a spectral autofluorescence correction method. The correction method is based on a simple algebraic operation using the fluorescence data from two detection channels and is applied on-the-fly in order to reliably identify hit beads by COPAS bead sorting. Our method provides a practical tool for the fast and efficient isolation of hit beads from one-bead one-compound library screens using either fluorescently labeled target proteins or biotinylated target proteins. This method makes hit bead identification easier and more reliable. It reduces false positives and eliminates the need for time-consuming pre-sorting of library beads in order to remove autofluorescent beads.
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