Development of a PNGase Rc Column for Online Deglycosylation of Complex Glycoproteins during HDX-MS

Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F86652036%3A_____%2F23%3A00581603" target="_blank" >RIV/86652036:_____/23:00581603 - isvavai.cz</a>
Alternative codes found
RIV/61388971:_____/23:00581603
Result on the web
<a href="https://pubs.acs.org/doi/10.1021/jasms.3c00268" target="_blank" >https://pubs.acs.org/doi/10.1021/jasms.3c00268</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1021/jasms.3c00268" target="_blank" >10.1021/jasms.3c00268</a>

Result language
angličtina
Original language name
Development of a PNGase Rc Column for Online Deglycosylation of Complex Glycoproteins during HDX-MS
Original language description
Protein glycosylation is one of the most common PTMs and many cell surface receptors, extracellular proteins, and biopharmaceuticals are glycosylated. However, HDX-MS analysis of such important glycoproteins has so far been limited by difficulties in determining the HDX of the protein segments that contain glycans. We have developed a column containing immobilized PNGase Rc (from Rudaea cellulosilytica) that can readily be implemented into a conventional HDX-MS setup to allow improved analysis of glycoproteins. We show that HDX-MS with the PNGase Rc column enables efficient online removal of N-linked glycans and the determination of the HDX of glycosylated regions in several complex glycoproteins. Additionally, we use the PNGase Rc column to perform a comprehensive HDX-MS mapping of the binding epitope of a mAb to c-Met, a complex glycoprotein drug target. Importantly, the column retains high activity in the presence of common quench-buffer additives like TCEP and urea and performed consistent across 114 days of extensive use. Overall, our work shows that HDX-MS with the integrated PNGase Rc column can enable fast and efficient online deglycosylation at harsh quench conditions to provide comprehensive analysis of complex glycoproteins.
Czech name
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Czech description
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Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology

Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year
2023
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

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