A hybrid monolithic column based on boronate-functionalized graphene oxide nanosheets for online specific enrichment of glycoproteins

Zhou, Chanyuan; Chen, Xiaoman; Du, Zuliang; Li, Gongke; Xiao, Xiaohua; Cai, Zongwei

doi:10.1016/j.chroma.2017.01.049

Cited by 44 publications

(23 citation statements)

References 35 publications

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“…Top-down MS/MS identification of intact fragments is more conclusive and convenient than traditional approaches using peptide map analysis of chromatography fractions 14 or SDS-PAGE bands. Several studies have recently used top-down, middle-down or middle-up MS/MS approaches to map the complete primary structure of a mAb, 19,22,23,[28][29][30] but these approaches have rarely been used to identify low level mAb fragments. Wang et al reported RP-HPLC HCD MS/MS characterization of the disulfide-reduced forms of an anti-C. difficile mAb and 7 product-related fragments (10-39 kDa MWs) fractionated by cation-exchange chromatography.…”

Section: Discussionmentioning

confidence: 99%

Specific and high-resolution identification of monoclonal antibody fragments detected by capillary electrophoresis–sodium dodecyl sulfate using reversed-phase HPLC with top-down mass spectrometry analysis

Wang

Cheung-Lau

Chen

et al. 2019

mAbs

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In recent years, capillary electrophoresis-sodium dodecyl sulfate (cSDS) has been widely used for high resolution separation and quantification of the fragments and aggregates of monoclonal antibodies (mAbs) to ensure the quality of mAb therapeutics. However, identification of the low-molecular-weight (LMW) and high-molecular-weight (HMW) species detected in cSDS electropherograms has been based primarily on the approximate MWs calculated from standard curves using known MW standards and correlations with fragments and aggregates identified by other methods. It is not easy to collect sufficient amounts of H/LMW species from cSDS for analysis by orthogonal methods and the direct coupling of cSDS with mass spectrometry (MS) is very difficult due to interference from SDS. In this study, we describe the precise identification of H/LMW species detected by cSDS using reversed-phase high performance liquid chromatography (RP-HPLC) coupled with top-down tandem MS analysis. The H/ LMW species were first identified by on-line RP-HPLC MS analysis and the RP-HPLC fractions were then analyzed by cSDS to connect the identified H/LMW species with the peaks in the cSDS electropherogram. With this method, 58 unique H/LMW species were identified from an immunoglobulin G1 (IgG1) mAb. The identified fragments ranged from 10 kDa single chain fragments to 130 kDa triple chain fragments, including some with post-translational modifications. This is the first study to clearly identify the antibody fragments, including the exact clipping sites, observed in cSDS electropherograms. The methodology and results presented here should be applicable to most other IgG1 mAbs.

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Section: Discussionmentioning

confidence: 99%

Specific and high-resolution identification of monoclonal antibody fragments detected by capillary electrophoresis–sodium dodecyl sulfate using reversed-phase HPLC with top-down mass spectrometry analysis

Wang

Cheung-Lau

Chen

et al. 2019

mAbs

View full text Add to dashboard Cite

show abstract

“…[145][146][147] Boronate affinity chromatography also has been of utility for enrichment of glycoproteins and glycopeptides, prior to mass spectrometric analysis. 148,149 With regard to size exclusion chromatography (SEC), excepting an investigation, no reports are available on its use for MD approach; Hummel et al have applied SEC to extract 2S albumin from the mustard. 138 Ion exchange chromatography, particularly weak cation exchange (WCX) chromatography in conjunction with hydrophilic interaction chromatography (HILIC) has been successfully applied for MD approach based studies (cf.…”

Section: Separation Technologymentioning

confidence: 99%

Middle-down approach: a choice to sequence and characterize proteins/proteomes by mass spectrometry

Pandeswari

Sabareesh

2019

RSC Adv.

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“…This MWNT-poly MMA-EDMA monolith has efficiently captured a 5000 fold diluted Hemoglobin from human blood samples. Recently, [37] they extended this work by replacing MWNT with boronate functionalized graphene oxide (GO) in MMA-EDMA monolith structure and applied it to successfully separate two glycoproteins (ovalbumin and ovotransferrin) from egg white. Incorporation of GO has tremendously increased the surface area of the monolith from 6.3 to 169.4 m 2 /g and without affecting the permeability.…”

Section: Gold Nanoparticles (Au Np)mentioning

confidence: 99%

Nanomaterial Fused Monolithic Microcolumns for Biomolecule Separation

Vijayalakshmi¹,

Tetala²

2018

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Achieving a highly pure biomolecule of interest is of great importance and chromatography based techniques are the reliable tools for this challenging task. During the last few decades, monoliths have been explored as alternative chromatography stationary phases for biomolecule separation. In this Minireview, we aim to provide information on different nanomaterials incorporated within the network of monoliths and strategies applied to perform nanomaterial surface functionalization. Subsequently, the applications of "nanomaterials embedded monolithic microcolumns" for separation of target biomolecules such as proteins and peptides with the help of different chromatography principles such as affinity, ionic and hydrophobic is also discussed in detail.

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A hybrid monolithic column based on boronate-functionalized graphene oxide nanosheets for online specific enrichment of glycoproteins

Cited by 44 publications

References 35 publications

Specific and high-resolution identification of monoclonal antibody fragments detected by capillary electrophoresis–sodium dodecyl sulfate using reversed-phase HPLC with top-down mass spectrometry analysis

Specific and high-resolution identification of monoclonal antibody fragments detected by capillary electrophoresis–sodium dodecyl sulfate using reversed-phase HPLC with top-down mass spectrometry analysis

Middle-down approach: a choice to sequence and characterize proteins/proteomes by mass spectrometry

Nanomaterial Fused Monolithic Microcolumns for Biomolecule Separation

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