High-throughput flow cytometric screening of combinatorial chemistry bead libraries for proteomics and drug discovery

James F. Leary; Lisa M. Reece; Xian-Bin Yang; David Gorenstein

doi:10.1117/12.589456

1 April 2005 High-throughput flow cytometric screening of combinatorial chemistry bead libraries for proteomics and drug discovery

James F. Leary, Lisa M. Reece, Xian-Bin Yang, David Gorenstein

Author Affiliations +

Proceedings Volume 5692, Advanced Biomedical and Clinical Diagnostic Systems III; (2005) https://doi.org/10.1117/12.589456
Event: SPIE BiOS, 2005, San Jose, CA, United States

Abstract

For proteomics drug discovery applications, combinatorial microbead thioaptamer libraries (one thioaptamer sequence per bead) are being created by split synthesis method, creating a "proteomics library" of protein capture beads which can be analyzed by high-throughput screening methods in this case, flow cytometry and cell sorting. Thioaptamers, oligonucleotides with thiophosphate backbone substitutions, function like antibodies in terms of recognizing specific protein sequences but have a number of advantages over antibody libraries. These proteomics beads can then be analyzed by high-speed flow cytometry and sorted to single-bead level depending on relative fluorescence brightness of fluorescently-labeled proteins, or for a specific protein from all of the molecules of cell subpopulations being analyzed. The thioaptamer sequences on a given bead showing high affinity for that protein can then be sequenced. Alternatively, the protein-capturing beads can be analyzed by MALDI-TOF mass spectrometry for analysis of the bound proteins. The beads can be thought of as equivalent to single-element positions of a proteomics chip arrays but with the advantage of being able to much more rapidly analyze hundreds of millions of possible amino acid sequences/epitopes on the basis of thioaptamer sequence affinities to select single sequences of interest. Additionally, those beads can be manipulated and isolated at the single bead level by high-throughput flow cytometry/cell sorting for subsequent sequencing of the thioaptamer sequences.

Citation Download Citation

James F. Leary, Lisa M. Reece, Xian-Bin Yang, and David Gorenstein "High-throughput flow cytometric screening of combinatorial chemistry bead libraries for proteomics and drug discovery", Proc. SPIE 5692, Advanced Biomedical and Clinical Diagnostic Systems III, (1 April 2005); https://doi.org/10.1117/12.589456

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