Proteomics

Dataset Information

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Engineered Peptide Barcodes for Binding Protein Generation


ABSTRACT: Binding protein generation relies on laborious screening cascades that process candidate molecules individually. To break with this paradigm, we developed NestLink, a binder selection and identification technology able to biophysically characterize thousands of library members at once without handling individual clones at any stage of the process. NestLink builds on genetically fused barcoding peptides, termed flycodes, which are designed for maximal detectability by mass spectrometry and serve as unique molecular identifiers for accurate deep sequencing. We applied NestLink to overcome current limitations of binder generation. Rare binders against an integral membrane protein were identified directly in the cellular environment of a human pathogen. Hundreds of binder candidates were simultaneously ranked according to kinetic parameters. Adverse effects of target immobilization were overcome by selecting nanobodies against an ABC transporter entirely in solution. NestLink may provide a basis for the selection of tailored binder characteristics directly in tissues or in living organisms

INSTRUMENT(S): Orbitrap Fusion, Q Exactive HF

ORGANISM(S): Synthetic Construct (code 12)

SUBMITTER: Bernd Roschitzki  

LAB HEAD: Prof. Markus Seeger

PROVIDER: PXD009301 | Pride | 2019-01-15

REPOSITORIES: Pride

Dataset's files

Source:
Action DRS
20160809_NL3idx6_expBtoC_06_B_BG.raw Raw
20160809_NL3idx6_expBtoC_09_B_MBP.raw Raw
20170303_04_aOMP_Ecoli1.raw Raw
20170303_05_aOMP_Ecoli2.raw Raw
20170303_06_aOMP_Ecoli3.raw Raw
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Binding protein generation typically relies on laborious screening cascades that process candidate molecules individually. We have developed NestLink, a binder selection and identification technology able to biophysically characterize thousands of library members at once without the need to handle individual clones at any stage of the process. NestLink uses genetically encoded barcoding peptides termed flycodes, which were designed for maximal detectability by mass spectrometry and support accur  ...[more]

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