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pH-switchable polymer that pulls down N-glycopeptides
A soluble poly(acrylic acid-co-hydrazide) captures glycopeptides in homogeneous solution and precipitates them by dropping pH for MS.
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What polymer enables homogeneous glycopeptide capture?
pH-responsive poly-(AA-co-hydrazide).
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
How fast/efficient is homogeneous enrichment claimed to be?
>95% efficiency within 1 h because mass transfer is unrestricted.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
What monomer ratio was used in the copolymer?
Methyl acrylate : acrylic acid molar ratio of 1 : 8.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
How are glycan diols activated for capture?
10 mM sodium periodate oxidizes them to aldehydes.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
What MALDI enhancement is quoted for m/z 1741.8?
Intensity ×29 and S/N ×325 after enrichment.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
How many non-redundant N-glycopeptides/proteins from mouse brain?
1317 glycopeptides and 458 glycoproteins.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
What reproducibility was reported across replicates?
80.0% of N-glycoproteins seen in at least two replicates.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
What false-discovery rate was estimated?
21 false N-glycopeptides; 1.46% average FDR.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
How is asialofetuin recovery described?
About 90% sample recovery by SDS-PAGE after release.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
How are N-glycosites localized after PNGase F?
A 0.984 Da mass shift at Asn on an N-X-S/T sequon.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified
How is the polymer cleaned before use?
Three low-pH-precipitation / high-pH-dissolution washes to remove poorly pH-responsive chains.
Source: A pH-responsive soluble polymer-based homogeneous system for fast and highly efficient <i>N</i>-glycoprotein/glycopeptide enrichment and identification by mass spectrometry · DOI · quote verified