Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry

Yudong Guan; Min Zhang; Jigang Wang; Hartmut Schlüter

doi:10.1021/acs.jproteome.1c00135

Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry

Standard

Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry. / Guan, Yudong; Zhang, Min; Wang, Jigang; Schlüter, Hartmut.

In: J PROTEOME RES, Vol. 20, No. 5, 07.05.2021, p. 2914-2922.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Guan, Y, Zhang, M, Wang, J & Schlüter, H 2021, 'Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry', J PROTEOME RES, vol. 20, no. 5, pp. 2914-2922. https://doi.org/10.1021/acs.jproteome.1c00135

APA

Guan, Y., Zhang, M., Wang, J., & Schlüter, H. (2021). Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry. J PROTEOME RES, 20(5), 2914-2922. https://doi.org/10.1021/acs.jproteome.1c00135

Vancouver

Guan Y, Zhang M, Wang J, Schlüter H. Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry. J PROTEOME RES. 2021 May 7;20(5):2914-2922. https://doi.org/10.1021/acs.jproteome.1c00135

Bibtex

@article{d55e2b6b88f14886bae73455d75675c3,

title = "Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry",

abstract = "Protein N-glycosylation characterization is challenging due to structural micro- and macro-heterogeneity. Although various N-glycan preparation strategies, including purification and derivatization, have been previously developed prior to mass spectrometric analysis, systematic evaluation still needs to be performed. This study compared the different N-glycan purification strategies, including filter-aided sample preparation, de-N-glycosylated protein precipitation, and trypsin digestion followed by reversed phase-based solid-phase extraction, and derivatization approaches, such as solid-phase permethylation, reductive amination, and reduction. With the comparative analysis, an optimized solid-phase permethylation (OSPP) workflow was developed for mass spectrometric N-glycomics, showing simplified analysis for N-glycan compositions and high yields using etanercept. The N-glycan samples released from trastuzumab and adalimumab were utilized to test OSPP to obtain their N-glycan profiles using mass spectrometry. Based on different standard procedures across laboratories, this study provides the reference for analysts to select an appropriate N-glycan preparation method with their research purposes.",

author = "Yudong Guan and Min Zhang and Jigang Wang and Hartmut Schl{\"u}ter",

year = "2021",

month = may,

day = "7",

doi = "10.1021/acs.jproteome.1c00135",

language = "English",

volume = "20",

pages = "2914--2922",

journal = "J PROTEOME RES",

issn = "1535-3893",

publisher = "American Chemical Society",

number = "5",

}

RIS

TY - JOUR

T1 - Comparative Analysis of Different N-glycan Preparation Approaches and Development of Optimized Solid-Phase Permethylation Using Mass Spectrometry

AU - Guan, Yudong

AU - Zhang, Min

AU - Wang, Jigang

AU - Schlüter, Hartmut

PY - 2021/5/7

Y1 - 2021/5/7

N2 - Protein N-glycosylation characterization is challenging due to structural micro- and macro-heterogeneity. Although various N-glycan preparation strategies, including purification and derivatization, have been previously developed prior to mass spectrometric analysis, systematic evaluation still needs to be performed. This study compared the different N-glycan purification strategies, including filter-aided sample preparation, de-N-glycosylated protein precipitation, and trypsin digestion followed by reversed phase-based solid-phase extraction, and derivatization approaches, such as solid-phase permethylation, reductive amination, and reduction. With the comparative analysis, an optimized solid-phase permethylation (OSPP) workflow was developed for mass spectrometric N-glycomics, showing simplified analysis for N-glycan compositions and high yields using etanercept. The N-glycan samples released from trastuzumab and adalimumab were utilized to test OSPP to obtain their N-glycan profiles using mass spectrometry. Based on different standard procedures across laboratories, this study provides the reference for analysts to select an appropriate N-glycan preparation method with their research purposes.

AB - Protein N-glycosylation characterization is challenging due to structural micro- and macro-heterogeneity. Although various N-glycan preparation strategies, including purification and derivatization, have been previously developed prior to mass spectrometric analysis, systematic evaluation still needs to be performed. This study compared the different N-glycan purification strategies, including filter-aided sample preparation, de-N-glycosylated protein precipitation, and trypsin digestion followed by reversed phase-based solid-phase extraction, and derivatization approaches, such as solid-phase permethylation, reductive amination, and reduction. With the comparative analysis, an optimized solid-phase permethylation (OSPP) workflow was developed for mass spectrometric N-glycomics, showing simplified analysis for N-glycan compositions and high yields using etanercept. The N-glycan samples released from trastuzumab and adalimumab were utilized to test OSPP to obtain their N-glycan profiles using mass spectrometry. Based on different standard procedures across laboratories, this study provides the reference for analysts to select an appropriate N-glycan preparation method with their research purposes.

U2 - 10.1021/acs.jproteome.1c00135

DO - 10.1021/acs.jproteome.1c00135

M3 - SCORING: Journal article

C2 - 33829797

VL - 20

SP - 2914

EP - 2922

JO - J PROTEOME RES

JF - J PROTEOME RES

SN - 1535-3893

IS - 5

ER -