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Multiplexed Competitive Screening of One-Bead-One-Component Combinatorial Libraries Using a ClonePix 2 Colony Sorter
R Ashton Lavoie1, Alice di Fazio2, Ruben G Carbonell3,4,5
1Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA. rabradl3@ncsu.edu.
International Journal of Molecular Sciences
|October 19, 2019
Summary
This study introduces a new method for screening peptide libraries to find selective binding ligands. It uses orthogonal fluorescent labeling and a ClonePix 2 Colony Picker to efficiently identify high-affinity binders for drug discovery applications.
Area of Science:
- Biochemistry
- Biotechnology
- Drug Discovery
Background:
- Solid-phase combinatorial library screening is crucial for identifying bioactive compounds.
- Ensuring binding selectivity of selected leads often lacks comprehensive strategies beyond basic blocking and competitive screening.
Purpose of the Study:
- To present a novel method for multiplexed solid-phase peptide library screening.
- To enhance the selection of high-affinity and selective ligands using orthogonal fluorescent labeling and advanced tracking.
Main Methods:
- Utilized a ClonePix 2 Colony Picker for multiplexed screening of peptide libraries.
- Integrated orthogonal fluorescent labeling for simultaneous positive and negative selection.
- Employed semi-quantitative tracking of target and competitor binding for individual library beads.
Main Results:
- Successfully screened a peptide library against green-labeled human immunoglobulin G (IgG) and red-labeled host cell proteins (HCPs).
- Identified 79 peptide ligand candidates (6.6%) as potential HCP-selective ligands.
- Demonstrated a screening rate exceeding 3,000 library beads per hour.
Conclusions:
- The presented method offers a robust strategy for selecting selective ligands with high affinity.
- This approach significantly improves the efficiency and comprehensiveness of solid-phase library screening.
- The technology is applicable for developing ligands for applications such as flow-through chromatography.
