Universal Solid-Phase Reversible Sample-Prep for Concurrent Proteome and N-Glycome Characterization

Journal of Proteome Research
Hui ZhouRichard S Lee

Abstract

We describe a novel solid-phase reversible sample-prep (SRS) platform that enables rapid sample preparation for concurrent proteome and N-glycome characterization for nearly all protein samples. SRS utilizes a uniquely functionalized, silica-based bead that has strong affinity toward proteins with minimal to no affinity for peptides and other small molecules. By leveraging this inherent size difference between proteins and peptides, SRS permits high-capacity binding of proteins, rapid removal of small molecules (detergents, metabolites, salts, peptides, etc.), extensive manipulation including enzymatic and chemical treatments on bead-bound proteins, and easy recovery of N-glycans and peptides. SRS was evaluated in a wide range of samples including glycoproteins, cell lysate, murine tissues, and human urine. SRS was also coupled to a quantitative strategy to investigate the differences between DU145 prostate cancer cells and its DIAPH3-silenced counterpart. Previous studies suggested that DIAPH3 silencing in DU145 induced transition to an amoeboid phenotype that correlated with tumor progression and metastasis. In this pilot study we identified distinct proteomic and N-glycomic alterations between them. A metastasis-associated t...Continue Reading

References

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Apr 21, 2012·Analytical Biochemistry·Hui ZhouRichard S Lee
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Citations

May 11, 2018·Journal of the American Society for Mass Spectrometry·John D BenktanderMilos V Novotny
Jan 21, 2017·Analytical Chemistry·Stefan GaunitzMilos V Novotny

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