Hydrogel-Encapsulated Beads Enable Proximity-Driven Encoded Library Synthesis and Screening

被引：4

作者：

Cavett, Valerie ^{[1
]}

Chan, Alix, I ^{[2
]}

Cunningham, Christian N. ^{[2
]}

Paegel, Brian M. ^{[1
,3
,4
]}

机构：

[1] Univ Calif Irvine, Dept Pharmaceut Sci, Irvine, CA 92697 USA

[2] Genentech Inc, Dept Peptide Therapeut, South San Francisco, CA 94080 USA

[3] Univ Calif Irvine, Dept Chem, Irvine, CA 92697 USA

[4] Univ Calif Irvine, Dept Biomed Engn, Irvine, CA 92697 USA

来源：

ACS CENTRAL SCIENCE | 2023年 / 9卷 / 08期

基金：

美国国家卫生研究院;

关键词：

SELECTION; DESIGN; PERMEABILITY; ACIDS;

D O I：

10.1021/acscentsci.3c00316

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Encoded combinatorial library technologies have dramaticallyexpandedthe chemical space for screening but are usually only analyzed byaffinity selection binding. It would be highly advantageous to reformatselection outputs to "one-bead-one-compound" solid-phaselibraries, unlocking activity-based and cellular screening capabilities.Here, we describe hydrogel-encapsulated magnetic beads that enablesuch a transformation. Bulk emulsion polymerization of polyacrylamidehydrogel shells around magnetic microbeads yielded uniform particles(7 & PLUSMN; 2 & mu;m diameter) that are compatible with diverse in-gelfunctionalization (amine, alkyne, oligonucleotides) and transformationsassociated with DNA-encoded library synthesis (acylation, enzymaticDNA ligation). In a case study of reformatting mRNA display libraries,transcription from DNA-templated magnetic beads encapsulated in gelparticles colocalized both RNA synthesis via hybridization with copolymerizedcomplementary DNA and translation via puromycin labeling. Two controlepitope templates (V5, HA) were successfully enriched (50- and 99-fold,respectively) from an NNK5 library bead screen via FACS.Proximity-driven library synthesis in concert with magnetic samplemanipulation provides a plausible means for reformatting encoded combinatoriallibraries at scale. Bead-templatedgel particles via emulsion polymerizationcan scalably transform display-type encoded libraries into one-bead-one-compoundlibraries suitable for functional screening.

引用

页码：1603 / 1610

页数：8

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