Simplifying Glycan Profiling through a High-Throughput Micropermethylation Strategy

Glycoproteins play key roles in various molecular and cellular functions and are among the most difficult to analyze biomolecules on account of their microheterogeneity, non-template-driven synthesis, and low abundances. The stability, serum half-life, immunogenicity, and biological activity of therapeutic glycoproteins, including antibodies, vaccines, and biomarkers, are regulated by their glycosylation profile. Thus, there is increasing demand for the qualitative and quantitative characterization and validation of glycosylation on glycoproteins. One of the most important derivatization processes for the structural characterization of released glycans by mass spectrometry (MS) is permethylation. We have recently developed a permethylation strategy in microscale that allows facile permethylation of glycans and permits the processing of large sample sets in nanogram amounts through high-throughput sample handling. Here, we are reporting the wide potential of micropermethylation-based high-throughput structural analysis of glycans from various sources, including human plasma, mammalian cells, and purified glycoproteins, through an automated tandem electrospray ionization–mass spectrometry (ESI-MSn) platform. The glycans released from the plasma, cells, and glycoproteins are permethylated in microscale in a 96-well plate or microcentrifuge tube and isolated by a C18 tip-based cleanup through a shorter and simple process. We have developed a workflow to accomplish an in-depth automated structural characterization MS program for permethylated N/O-glycans through an automated high-throughput multistage tandem MS acquisition. We have demonstrated the utility of this workflow using the examples of sialic acid linkages and bisecting GlcNAc ( N-acetylglucosamine) on the glycans. This approach can automate the high-throughput screening of glycosylation on large sample sets of glycoproteins, including clinical glycan biomarkers and glycoprotein therapeutics.

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Evaluation of ambient mass spectrometry tools for assessing inherent postharvest pepper quality

Horticulture Research ◽

10.1038/s41438-021-00596-x ◽

2021 ◽

Vol 8 (1) ◽

Author(s):

Tyler J. Mason ◽

Harmonie M. Bettenhausen ◽

Jacqueline M. Chaparro ◽

Mark E. Uchanski ◽

Jessica E. Prenni

Keyword(s):

Mass Spectrometry ◽

High Throughput ◽

High Throughput Screening ◽

Direct Analysis ◽

Quality Characteristics ◽

Ambient Mass Spectrometry ◽

Postharvest Quality ◽

Ionization Mass ◽

Ambient Conditions ◽

Assess Quality

AbstractHorticulturists are interested in evaluating how cultivar, environment, or production system inputs can affect postharvest quality. Ambient mass spectrometry approaches enable analysis of minimally processed samples under ambient conditions and offer an attractive high-throughput alternative for assessing quality characteristics in plant products. Here, we evaluate direct analysis in real time (DART-MS) mass spectrometry and rapid evaporative ionization-mass spectrometry (REIMS) to assess quality characteristics in various pepper (Capsicum annuum L.) cultivars. DART-MS exhibited the ability to discriminate between pod colors and pungency based on chemical fingerprints, while REIMS could distinguish pepper market class (e.g., bell, lunchbox, and popper). Furthermore, DART-MS analysis resulted in the putative detection of important bioactive compounds in human diet such as vitamin C, p-coumaric acid, and capsaicin. The results of this study demonstrate the potential for these approaches as accessible and reliable tools for high throughput screening of pepper quality.

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MALDIViz: A Comprehensive Informatics Tool for MALDI-MS Data Visualization and Analysis

SLAS DISCOVERY Advancing Life Sciences ◽

10.1177/2472555217727517 ◽

2017 ◽

Vol 22 (10) ◽

pp. 1246-1252 ◽

Cited By ~ 1

Author(s):

Kishore Kumar Jagadeesan ◽

Simon Ekström

Keyword(s):

Mass Spectrometry ◽

High Throughput ◽

High Throughput Screening ◽

Web Application ◽

Low Cost ◽

Maldi Ms ◽

Ionization Mass ◽

Label Free ◽

Label Free Detection ◽

R Shiny

Recently, mass spectrometry (MS) has emerged as an important tool for high-throughput screening (HTS) providing a direct and label-free detection method, complementing traditional fluorescent and colorimetric methodologies. Among the various MS techniques used for HTS, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) provides many of the characteristics required for high-throughput analyses, such as low cost, speed, and automation. However, visualization and analysis of the large datasets generated by HTS MALDI-MS can pose significant challenges, especially for multiparametric experiments. The datasets can be generated fast, and the complexity of the experimental data (e.g., screening many different sorbent phases, the sorbent mass, and the load, wash, and elution conditions) makes manual data analysis difficult. To address these challenges, a comprehensive informatics tool called MALDIViz was developed. This tool is an R-Shiny-based web application, accessible independently of the operating system and without the need to install any program locally. It has been designed to facilitate easy analysis and visualization of MALDI-MS datasets, comparison of multiplex experiments, and export of the analysis results to high-quality images.

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A simpler sampling interface of venturi easy ambient sonic-spray ionization mass spectrometry for high-throughput screening enzyme inhibitors

Analytica Chimica Acta ◽

10.1016/j.aca.2016.01.051 ◽

2016 ◽

Vol 913 ◽

pp. 86-93 ◽

Cited By ~ 5

Author(s):

Ning Liu ◽

Yang Liu ◽

YuHan Yang ◽

Lan He ◽

Jin Ouyang

Keyword(s):

Mass Spectrometry ◽

High Throughput ◽

High Throughput Screening ◽

Enzyme Inhibitors ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Sonic Spray Ionization ◽

Sampling Interface

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Flow injection analysis electrospray ionization mass spectrometry for high-throughput screening of a gene library for amidase activity

Analytical Biochemistry ◽

10.1016/j.ab.2009.07.018 ◽

2009 ◽

Vol 394 (2) ◽

pp. 159-163 ◽

Cited By ~ 3

Author(s):

Huub J.W. Henderickx ◽

Petronella C. Raemakers-Franken ◽

Sjoerd van der Wal ◽

Chris G. de Koster ◽

Alexander L.L. Duchateau ◽

...

Keyword(s):

Mass Spectrometry ◽

Electrospray Ionization ◽

High Throughput ◽

Flow Injection ◽

Flow Injection Analysis ◽

Electrospray Ionization Mass Spectrometry ◽

High Throughput Screening ◽

Ionization Mass ◽

Gene Library ◽

Amidase Activity

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Fully automated chip-based nanoelectrospray ionization-mass spectrometry as an effective tool for rapid and high-throughput screening of 5α-reductase inhibitors

Analytical and Bioanalytical Chemistry ◽

10.1007/s00216-020-02408-y ◽

2020 ◽

Vol 412 (7) ◽

pp. 1685-1692 ◽

Cited By ~ 1

Author(s):

Ruirong Zheng ◽

Linnan Li ◽

Xueli Deng ◽

Mei Tian ◽

Zhengtao Wang ◽

...

Keyword(s):

Mass Spectrometry ◽

High Throughput ◽

High Throughput Screening ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Reductase Inhibitors ◽

Nanoelectrospray Ionization

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High-throughput screening of organic reactions in microdroplets using desorption electrospray ionization mass spectrometry (DESI-MS): hardware and software implementation

Analytical Methods ◽

10.1039/d0ay00072h ◽

2020 ◽

Vol 12 (28) ◽

pp. 3654-3669

Author(s):

Tiago Jose P. Sobreira ◽

Larisa Avramova ◽

Botond Szilagyi ◽

David L. Logsdon ◽

Bradley P. Loren ◽

...

Keyword(s):

Mass Spectrometry ◽

Electrospray Ionization ◽

High Throughput ◽

Electrospray Ionization Mass Spectrometry ◽

High Throughput Screening ◽

Desorption Electrospray Ionization ◽

Ionization Mass Spectrometry ◽

Ionization Mass ◽

Desi Ms ◽

Novel Method

Implementation of a novel method for high-throughput screening of reactions in microdroplets. The reaction and analysis steps are performed simultaneously using desorption electrospray ionization mass spectrometry (DESI-MS) at a rate of up to 1 reaction mixture per second.

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