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Monitoring the glycosylation status of proteins using Raman spectroscopy

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Monitoring the glycosylation status of proteins using Raman spectroscopy. / Brewster, Victoria L.; Ashton, Lorna; Goodacre, Royston.

In: Analytical Chemistry, Vol. 83, No. 15, 01.08.2011, p. 6074-6081.

Research output: Contribution to journalJournal articlepeer-review

Harvard

Brewster, VL, Ashton, L & Goodacre, R 2011, 'Monitoring the glycosylation status of proteins using Raman spectroscopy', Analytical Chemistry, vol. 83, no. 15, pp. 6074-6081. https://doi.org/10.1021/ac2012009

APA

Brewster, V. L., Ashton, L., & Goodacre, R. (2011). Monitoring the glycosylation status of proteins using Raman spectroscopy. Analytical Chemistry, 83(15), 6074-6081. https://doi.org/10.1021/ac2012009

Vancouver

Brewster VL, Ashton L, Goodacre R. Monitoring the glycosylation status of proteins using Raman spectroscopy. Analytical Chemistry. 2011 Aug 1;83(15):6074-6081. https://doi.org/10.1021/ac2012009

Author

Brewster, Victoria L. ; Ashton, Lorna ; Goodacre, Royston. / Monitoring the glycosylation status of proteins using Raman spectroscopy. In: Analytical Chemistry. 2011 ; Vol. 83, No. 15. pp. 6074-6081.

Bibtex

@article{9d6450fc1ccd4de98b6465421da42911,
title = "Monitoring the glycosylation status of proteins using Raman spectroscopy",
abstract = "Protein-based biopharmaceuticals are becoming increasingly widely used as therapeutic agents, and the characterization of these biopharmaceuticals poses a significant analytical challenge. In particular, monitoring posttranslational modifications (PTMs), such as glycosylation, is an important aspect of this characterization because these glycans can strongly affect the stability, immunogenicity, and pharmacolcinetics of these biotherapeutic drugs. Raman spectroscopy is a powerful tool, with many emerging applications in the bioprocessing arena. Although the technique has a relatively rich history in protein science, only recently has Raman spectroscopy been investigated for assessing posttranslational modifications, including phosphorylation, acetylation, trimethylation, and ubiquitination. In this investigation, we develop for the first time Raman spectroscopy combined with multivariate data analyses, including principal components analysis and partial least-squares regression, for the determination of the glycosylation status of proteins and quantifying the relative concentrations of the native ribonuclease (RNase) A protein and RNase B glycoprotein within mixtures.",
keywords = "OPTICAL-ACTIVITY, TRIFLUOROMETHANESULFONIC ACID, 2-DIMENSIONAL RAMAN, LINKED GLYCANS, DEGLYCOSYLATION, GLYCOPROTEINS, QUANTIFICATION, IDENTIFICATION, CONFORMATION, POLYPEPTIDE",
author = "Brewster, {Victoria L.} and Lorna Ashton and Royston Goodacre",
year = "2011",
month = aug,
day = "1",
doi = "10.1021/ac2012009",
language = "English",
volume = "83",
pages = "6074--6081",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "American Chemical Society",
number = "15",

}

RIS

TY - JOUR

T1 - Monitoring the glycosylation status of proteins using Raman spectroscopy

AU - Brewster, Victoria L.

AU - Ashton, Lorna

AU - Goodacre, Royston

PY - 2011/8/1

Y1 - 2011/8/1

N2 - Protein-based biopharmaceuticals are becoming increasingly widely used as therapeutic agents, and the characterization of these biopharmaceuticals poses a significant analytical challenge. In particular, monitoring posttranslational modifications (PTMs), such as glycosylation, is an important aspect of this characterization because these glycans can strongly affect the stability, immunogenicity, and pharmacolcinetics of these biotherapeutic drugs. Raman spectroscopy is a powerful tool, with many emerging applications in the bioprocessing arena. Although the technique has a relatively rich history in protein science, only recently has Raman spectroscopy been investigated for assessing posttranslational modifications, including phosphorylation, acetylation, trimethylation, and ubiquitination. In this investigation, we develop for the first time Raman spectroscopy combined with multivariate data analyses, including principal components analysis and partial least-squares regression, for the determination of the glycosylation status of proteins and quantifying the relative concentrations of the native ribonuclease (RNase) A protein and RNase B glycoprotein within mixtures.

AB - Protein-based biopharmaceuticals are becoming increasingly widely used as therapeutic agents, and the characterization of these biopharmaceuticals poses a significant analytical challenge. In particular, monitoring posttranslational modifications (PTMs), such as glycosylation, is an important aspect of this characterization because these glycans can strongly affect the stability, immunogenicity, and pharmacolcinetics of these biotherapeutic drugs. Raman spectroscopy is a powerful tool, with many emerging applications in the bioprocessing arena. Although the technique has a relatively rich history in protein science, only recently has Raman spectroscopy been investigated for assessing posttranslational modifications, including phosphorylation, acetylation, trimethylation, and ubiquitination. In this investigation, we develop for the first time Raman spectroscopy combined with multivariate data analyses, including principal components analysis and partial least-squares regression, for the determination of the glycosylation status of proteins and quantifying the relative concentrations of the native ribonuclease (RNase) A protein and RNase B glycoprotein within mixtures.

KW - OPTICAL-ACTIVITY

KW - TRIFLUOROMETHANESULFONIC ACID

KW - 2-DIMENSIONAL RAMAN

KW - LINKED GLYCANS

KW - DEGLYCOSYLATION

KW - GLYCOPROTEINS

KW - QUANTIFICATION

KW - IDENTIFICATION

KW - CONFORMATION

KW - POLYPEPTIDE

U2 - 10.1021/ac2012009

DO - 10.1021/ac2012009

M3 - Journal article

VL - 83

SP - 6074

EP - 6081

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 15

ER -