Top-down proteomics

Nature Reviews Methods Primers, Journal Year: 2024, Volume and Issue: 4(1)

Published: June 13, 2024

Language: Английский

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Recent Advances in Mass Spectrometry-Based Bottom-Up Proteomics

Analytical Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

Mass spectrometry-based proteomics is about 35 years old, and recent progress appears to be speeding up across all subfields. In this review, we focus on advances over the last two in select areas within bottom-up proteomics, including approaches high-throughput experiments, data analysis using machine learning, drug discovery, glycoproteomics, extracellular vesicle structural proteomics.

Language: Английский

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Impact of Excipient Extraction and Buffer Exchange on Recombinant Monoclonal Antibody Stability

Molecular Pharmaceutics, Journal Year: 2024, Volume and Issue: 21(4), P. 1872 - 1883

Published: Feb. 29, 2024

The foundation of a biosimilar manufacturer's regulatory filing is the demonstration analytical and functional similarity between product pertinent originator product. excipients in formulation may interfere with characterization using typical techniques during this biosimilarity exercise. Consequently, producers products resort to buffer exchange isolate biotherapeutic protein from drug formulation. However, impact that isolation has on stability not completely known. This study aims elucidate extent which mAb via ultrafiltration-diafiltration-based impacts stability. It been demonstrated repeated extraction cycles do result significant changes higher-order structure (red-shift 5.0 nm fluorescence maxima exchanged samples) also an increase formation basic variants 19.1 26.7% 32.3 36.9% extracted innovator Tmab samples, respectively. was observed under certain conditions tertiary disruptions, could be restabilized depending composition. Thus, through Based observations reported paper, we recommend manufacturers take into consideration these effects when performing assessments.

Language: Английский

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Quality control of allergen products with mass spectrometry part I: Positioning within the EU regulatory framework

Allergy, Journal Year: 2024, Volume and Issue: 79(8), P. 2088 - 2096

Published: Feb. 29, 2024

Abstract Mass spectrometry (MS) has advanced greatly and many of its applications are ready for utilization within regulatory procedures could significantly contribute to overcome challenges in standardization allergen products. It seems sensible discuss MS the framework, before addressing technical questions. While application purified proteins is well established from product development manufacturer's release analytics, complex products such as still under development. needs be determined where it can complement or replace methods offers limited improvement. Despite appeal versatility, currently mentioned guidelines only one possible measurement method. For example, no specific method given European Pharmacopoeia. We EU framework. This includes their advantages disadvantages positioning between research, characterization, analytics official batch testing. qualitative detection single multiple allergens proof identity, semi‐quantitative protein profiles consistency testing, quantification state mass units allergens. may also facilitate products, reference standards.

Language: Английский

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Ion Mobility-Mass Spectrometry and Collision-Induced Unfolding Rapidly Characterize the Structural Polydispersity and Stability of an Fc-Fusion Protein

Analytical Chemistry, Journal Year: 2024, Volume and Issue: 96(24), P. 10003 - 10012

Published: June 10, 2024

Fc-fusion proteins are an emerging class of protein therapeutics that combine the properties biological ligands with unique fragment crystallizable (Fc) domain immunoglobulin G (IgG). Due to their diverse higher-order structures (HOSs), remain challenging characterization targets within biopharmaceutical pipelines. While high-resolution biophysical tools available for HOS characterization, they frequently demand extended time frames and substantial quantities purified samples, rendering them impractical swiftly screening candidate molecules. Herein, we describe development ion mobility-mass spectrometry (IM-MS) collision-induced unfolding (CIU) workflows aim fill this technology gap, where focus on probing a model Fc-Interleukin-10 (Fc-IL-10) fusion engineered using flexible glycine-serine linkers. We evaluate ability these techniques probe flexibility Fc-IL-10 in absence bulk solvent relative other similar size, as well localize structural changes low charge state ions specific Fc IL-10 events during CIU. subsequently apply local effects linkers stability homodimer, which is biologically active form IL-10. Our data reveals produces significantly more transitions CIU broader IM profiles when compared wide range proteins, indicative its exceptional dynamism. Furthermore, use combination enzymatic approaches annotate intricate domains Fc-IL-10. Finally, detect strong positive, quadratic relationship between average linker mass stability, suggesting cooperative influence overall stability. This first reported study IM-MS characterize illustrating practical applicability approach.

Language: Английский

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Biosimilars in the Era of Artificial Intelligence—International Regulations and the Use in Oncological Treatments

Pharmaceuticals, Journal Year: 2024, Volume and Issue: 17(7), P. 925 - 925

Published: July 10, 2024

This research is based on three fundamental aspects of successful biosimilar development in the challenging biopharmaceutical market. First, regulations eight selected countries: Japan, South Korea, United States, Canada, Brazil, Argentina, Australia, and Africa, represent four continents. The regulatory countries studied are analyzed, highlighting challenges facing biosimilars, including their complex approval processes need for standardized guidelines. There an inconsistency depending whether used a developed or developing country. In observed, biosimilars considered excellent alternatives to patent-protected biological products treatment chronic diseases. second aspect addressed, various analytical AI modeling methods (such as machine learning tools, reinforcement learning, supervised, unsupervised, deep tools) were analyzed observe patterns that lead prevalence cancer model behaviors most prominent active compounds with spectroscopy. Finally, analysis use approved by FDA EMA was proposed.

Language: Английский

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Characterization of Higher Order Structural Changes of a Thermally Stressed Monoclonal Antibody via Mass Spectrometry Footprinting and Other Biophysical Approaches

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(46), P. 16840 - 16849

Published: Nov. 7, 2023

Characterizing changes in the higher order structure (HOS) of monoclonal antibodies upon stressed conditions is critical to gaining a better understanding product and process. One single biophysical approach may not be best suited assess HOS comprehensively; thus, synergy from multiple, complementary approaches improves characterization accuracy resolution. In this study, we employed two mass spectrometry (MS )-based footprinting techniques, namely, fast photochemical oxidation proteins (FPOP)-MS hydrogen–deuterium exchange (HDX)-MS, supported by dynamic light scattering (DLS), differential scanning calorimetry (DSC), circular dichroism (CD), nuclear magnetic resonance (NMR) study mAb thermal stress. The techniques report nuanced which CD detects no secondary or tertiary structure, yet DLS measurements show an increase hydrodynamic radius. DSC indicates that stability decreases, chemical conformational accumulate with incubation time according NMR. Furthermore, whereas HDX-MS does indicate changes, FPOP-MS reveals at residue resolution for some amino acids. local phenomena observed several residues various patterns degradation during stress: change, solvent exposure, biphasic response exposure. All evidences efficiently resolves subtle structural novel pathways stress treatment residue-level

Language: Английский

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Exploring the structural heterogeneity of IgG1s: A comprehensive analysis of glycosylated and deglycosylated forms using TIMS-TOF-MS technique

Microchemical Journal, Journal Year: 2024, Volume and Issue: 207, P. 112099 - 112099

Published: Nov. 3, 2024

Language: Английский

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