PXD026359
PXD026359 is an original dataset announced via ProteomeXchange.
Dataset Summary
| Title | Peptide barcoding for one-pot evaluation of sequence-function relationships of free nanobodies |
| Description | Optimization of protein binders relies on laborious screening processes. Particularly slow is investigation of sequence-function relationships of protein binders in which diverse amino-acid-substituted mutants are constructed, purified, and evaluated individually. Here, we developed peptide barcoding 2.0, a high-throughput approach for accurate investigation of sequence-function relationships of hundreds of protein binders at once. Our approach is based on combining generation of a mutagenized protein binder library fused with unique peptide barcodes, formation of binder–antigen complexes at different ratios, fine fractionation of binder–antigen complexes by size exclusion chromatography, and highly specific quantification of peptide barcodes by mass spectrometry. Applying peptide barcoding 2.0 for evaluation of anti-green fluorescent protein nanobody as a model, we succeeded in identifying various mutant Nbs with decreased affinities at once. Moreover, we successfully discriminated subtle changes in KD at the order of nM to sub-nM. These results showed that peptide barcoding 2.0 is a powerful tool for engineering of protein binders, enabling reliable one-pot evaluation of sequence-function relationships. |
| HostingRepository | jPOST |
| AnnounceDate | 2021-05-30 |
| AnnouncementXML | Submission_2022-09-18_03:43:53.921.xml |
| DigitalObjectIdentifier | |
| ReviewLevel | Non peer-reviewed dataset |
| DatasetOrigin | Original dataset |
| RepositorySupport | Unsupported dataset by repository |
| PrimarySubmitter | Yusei Matsuzaki |
| SpeciesList | scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932; |
| ModificationList | S-carboxymethyl-L-cysteine; alpha-amino acetylated residue |
| Instrument | LCMS-8060 |
Dataset History
| Revision | Datetime | Status | ChangeLog Entry |
|---|---|---|---|
| 0 | 2021-05-29 21:29:50 | ID requested | |
| 1 | 2021-05-29 21:54:21 | announced | |
| ⏵ 2 | 2022-09-18 03:43:54 | announced | 2022-09-18: Updated FTP location. |
Publication List
| Dataset with its publication pending |
Keyword List
| submitter keyword: peptide barcoding, nanobody |
Contact List
| Wataru Aoki | |
|---|---|
| lab head | |
| Yusei Matsuzaki | |
| contact affiliation | Kyoto University |
| dataset submitter | |
Full Dataset Link List
| jPOST dataset URI |
| Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.jpostdb.org/JPST001198/ |
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