Assembly and use of high-density recombinant peptide chips for large-scale ligand screening is a practical alternative to synthetic peptide libraries

作者：Hund**erger Harald; Oender Kamil; Schuller Goetzburg Peter; Virok Dezso P; Herzog Julia; Rid Raphaela

来源：BMC Genomics, 2017, 18(1): 450.

DOI：10.1186/s12864-017-3814-3

• Peptide library
• Recombinational cloning
• Recombinant peptide array
• Protein chip
• Peptide screening
• Protein tags
• Ligand design
• High affinity
• Target binding
• Diversity

Background: Recombinant peptide chips could constitute a versatile complementation to state-of-the-art in situ (chemical on-chip) synthesis, particle-based printing, or pre-manufactured peptide spotting. Bottlenecks still impeding a routine implementation-from restricted peptide lengths, low diversity and low array densities to high costs-could so be overcome.
Methods: To assess overall performance, we assembled recombinant chips composed of 38,400 individual peptide spots on the area of a standard 96-well microtiter plate from comprehensive, highly diverse (>107 single clones) short random peptide libraries.
Results: Screening of altogether 476,160 clones against Streptavidin uncovered 2 discrete new binders: a characteristic HPQ-motif containing VSHPQAPF and a cyclic CSGSYGSC peptide. Interactions were technically confirmed by fluorescence polarization as well as biolayer-interferometry, and their potential suitability as novel detection tags evaluated by detection of a peptide-fused exemplary test protein.
Conclusion: From our data we conclude that the presented technical pipeline can reliably identify novel hits, useful as first-generation binders or templates for subsequent ligand design plus engineering.

• 出版日期2017-6-8