Exploring the Chemical Space of Protein Glycosylation in Noncovalent Protein Complexes: An Expedition along Different Structural Levels of Human Chorionic Gonadotropin by Employing Mass Spectrometry

Lebede, Maximilian; Marco, Fiammetta Di; Esser‐Skala, Wolfgang; Hennig, René; Wohlschlager, Therese; Huber, Christian G.

doi:10.1021/acs.analchem.1c02199

Cited by 12 publications

(19 citation statements)

References 47 publications

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“…In an elegant combination of glycoproteoform profiling and non-covalent mass spectrometry, Lebede et al used high-resolution native MS to analyze recombinant human chorionic gonadotropin (hCG, Ovitrelle). The protein hormone hCG is involved in early embryo–maternal communication and maintenance of pregnancy.…”

Section: Proteoform Profiling By High-resolution Native Msmentioning

confidence: 99%

High-Resolution Native Mass Spectrometry

2021

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Native mass spectrometry (MS) involves the analysis and characterization of macromolecules, predominantly intact proteins and protein complexes, whereby as much as possible the native structural features of the analytes are retained. As such, native MS enables the study of secondary, tertiary, and even quaternary structure of proteins and other biomolecules. Native MS represents a relatively recent addition to the analytical toolbox of mass spectrometry and has over the past decade experienced immense growth, especially in enhancing sensitivity and resolving power but also in ease of use. With the advent of dedicated mass analyzers, sample preparation and separation approaches, targeted fragmentation techniques, and software solutions, the number of practitioners and novel applications has risen in both academia and industry. This review focuses on recent developments, particularly in high-resolution native MS, describing applications in the structural analysis of protein assemblies, proteoform profiling ofamong othersbiopharmaceuticals and plasma proteins, and quantitative and qualitative analysis of protein–ligand interactions, with the latter covering lipid, drug, and carbohydrate molecules, to name a few.

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Section: Proteoform Profiling By High-resolution Native Msmentioning

confidence: 99%

High-Resolution Native Mass Spectrometry

2021

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“…For reduction of computing time, an initial global characterization of the Nglycome can be performed by CE. The resulting glycan list is used to search against a targeted set of defined N-glycan structures as variable protein/peptide modifications (Thaysen-Andersen and Packer 2014; Parker et al 2013;Lebede et al 2021;Pralow et al 2021;Pioch et al 2018). This pre-knowledge of attached glycan structures greatly reduces the search space, which in turn significantly decreases the computing time.…”

Section: Coupling Capillary Electrophoresis With Mass Spectrometrymentioning

confidence: 99%

Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis

Cajic

Hennig

Burock³

et al. 2021

Experientia Supplementum

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The in-depth characterization of protein glycosylation has become indispensable in many research fields and in the biopharmaceutical industry. Especially knowledge about modulations in immunoglobulin G (IgG) N-glycosylation and their effect on immunity enabled a better understanding of human diseases and the development of new, more effective drugs for their treatment. This chapter provides a deeper insight into capillary (gel) electrophoresis-based (C(G)E) glycan analysis, addressing its impressive performance and possibilities, its great potential regarding real high-throughput for large cohort studies, as well as its challenges and limitations. We focus on the latest developments with respect to miniaturization and mass spectrometry coupling, as well as data analysis and interpretation. The use of exoglycosidase sequencing in combination with current C(G)E technology is discussed, highlighting possible difficulties and pitfalls. The application section describes the detailed characterization of N-glycosylation, utilizing multiplexed CGE with laser-induced fluorescence detection (xCGE-LIF). Besides a comprehensive overview on antibody glycosylation by comparing species-specific IgGs and human immunoglobulins A, D, E, G, and M, the chapter comprises a comparison of therapeutic monoclonal antibodies from different production cell lines, as well as a detailed characterization of Fab and Fc glycosylation. These examples illustrate the full potential of C(G)E, resolving the smallest differences in sugar composition and structure.

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“… Graph representations of the linear experimental sections in van Faassen et al, 28 Lajin and Goessler, 29 and Kocurek et al 30 (left), ”parallel” experiments in McAvan et al, 31 Pérez-Diez et al, 32 and Yan et al 17 (middle), and more complex methods in Venzac et al, 33 Flint et al, 34 and Lebede et al 35 (right). The graphs were rendered and the nodes colored as in Figure 1 .…”

Section: Resultsmentioning

confidence: 99%

Semantic Annotation of Experimental Methods in Analytical Chemistry

et al. 2022

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A major obstacle for reusing and integrating existing data is finding the data that is most relevant in a given context. The primary metadata resource is the scientific literature describing the experiments that produced the data. To stimulate the development of natural language processing methods for extracting this information from articles, we have manually annotated 100 recent open access publications in Analytical Chemistry as semantic graphs. We focused on articles mentioning mass spectrometry in their experimental sections, as we are particularly interested in the topic, which is also within the domain of several ontologies and controlled vocabularies. The resulting gold standard dataset is publicly available and directly applicable to validating automated methods for retrieving this metadata from the literature. In the process, we also made a number of observations on the structure and description of experiments and open access publication in this journal.

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Exploring the Chemical Space of Protein Glycosylation in Noncovalent Protein Complexes: An Expedition along Different Structural Levels of Human Chorionic Gonadotropin by Employing Mass Spectrometry

Cited by 12 publications

References 47 publications

High-Resolution Native Mass Spectrometry

High-Resolution Native Mass Spectrometry

Capillary (Gel) Electrophoresis-Based Methods for Immunoglobulin (G) Glycosylation Analysis

Semantic Annotation of Experimental Methods in Analytical Chemistry

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