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Open AccessCommunication

Multiplexed Competitive Screening of One-Bead-One-Component Combinatorial Libraries Using a ClonePix 2 Colony Sorter

1
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC 27695, USA
2
Biomanufacturing Training and Education Center (BTEC), North Carolina State University, Raleigh, NC 27695, USA
3
National Institute for Innovation in Manufacturing Biopharmaceuticals (NIIMBL), Newark, DE 19711, USA
*
Authors to whom correspondence should be addressed.
Int. J. Mol. Sci. 2019, 20(20), 5119; https://doi.org/10.3390/ijms20205119
Received: 25 September 2019 / Revised: 12 October 2019 / Accepted: 13 October 2019 / Published: 16 October 2019
(This article belongs to the Collection Feature Papers in Molecular Biophysics)
Screening solid-phase combinatorial libraries of bioactive compounds against fluorescently labeled target biomolecules is an established technology in ligand and drug discovery. Rarely, however, do screening methods include comprehensive strategies—beyond mere library blocking and competitive screening—to ensure binding selectivity of selected leads. This work presents a method for multiplexed solid-phase peptide library screening using a ClonePix 2 Colony Picker that integrates (i) orthogonal fluorescent labeling for positive selection against a target protein and negative selection against competitor species with (ii) semi-quantitative tracking of target vs. competitor binding for every library bead. The ClonePix 2 technology enables global at-a-glance evaluation and customization of the parameters for bead selection to ensure high affinity and selectivity of the isolated leads. A case study is presented by screening a peptide library against green-labeled human immunoglobulin G (IgG) and red-labeled host cell proteins (HCPs) using ClonePix 2 to select HCP-binding ligands for flow-through chromatography applications. Using this approach, 79 peptide ligand candidates (6.6% of the total number of ligands screened) were identified as potential HCP-selective ligands, enabling a potential rate of >3,000 library beads screened per hour. View Full-Text
Keywords: library screening; bioactive peptides; ligand development; affinity chromatography; affinity ligand; ClonePix 2 library screening; bioactive peptides; ligand development; affinity chromatography; affinity ligand; ClonePix 2
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Lavoie, R.A.; di Fazio, A.; Carbonell, R.G.; Menegatti, S. Multiplexed Competitive Screening of One-Bead-One-Component Combinatorial Libraries Using a ClonePix 2 Colony Sorter. Int. J. Mol. Sci. 2019, 20, 5119.

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