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MALDI-TOF MS analysis for detection of bovine coronavirus with tryptic peptides from viral proteins.
J Microorg Control
January 2025
Division of Microbiology, National Institute of Health Sciences.
Bovine coronavirus (BCoV）, a significant cattle pathogen causing enteric and respiratory diseases, is primarily detected using reverse transcription-polymerase chain reaction. Our objective was to develop a novel detection method for BCoV by matrix-assisted laser desorption/ionization‒time-of-flight mass spectrometry (MALDI-TOF MS）. Peptide mass fingerprint analysis revealed that nucleocapsid (N）, membrane (M）, and hemagglutinin-esterase (HE) were three main BCoV proteins.
View Article and Find Full Text PDFThe Enzyme Effect: Broadening the Horizon of MS Optimization to Nontryptic Digestion in Proteomics.
J Am Soc Mass Spectrom
January 2025
MS Proteomics Research Group, HUN-REN Research Centre for Natural Sciences, Magyar Tudósok körútja 2, H-1117 Budapest, Hungary.
In recent years, alternative enzymes with varied specificities have gained importance in MS-based bottom-up proteomics, offering orthogonal information about biological samples and advantages in certain applications. However, most mass spectrometric workflows are optimized for tryptic digests. This raises the questions of whether enzyme specificity impacts mass spectrometry and if current methods for nontryptic digests are suboptimal.
View Article and Find Full Text PDFSimultaneous isotyping and semi-quantitation of anti-drug antibodies to an IgG1 biotherapeutic using hybrid LBA-LC-MS/MS.
Bioanalysis
December 2024
ICON Bioanalytical Laboratories, Assen, The Netherlands.
Background: Commonly, ligand-binding platforms are being used for immunogenicity assessment, but with the recent advent of liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS) for protein quantification, this technology has become an alternative for the measurement of anti-drug antibodies (ADAs), when combined with an immunocapture step to extract them out of the biological sample.
Method: The monoclonal antibody adalimumab was immobilized on magnetic beads to isolate ADAs against this drug from serum samples. Multiple repetitions of immunopurification were used to minimize nonspecific binding and improve drug tolerance while maintaining sufficient recovery.
Simple endoglycosidase-assisted peptide mapping workflow for characterizing non-consensus n-glycosylation in therapeutic monoclonal antibodies.
J Pharm Sci
November 2024
Analytical Chemistry, Regeneron Pharmaceuticals Inc., Tarrytown, NY, USA.
Article Synopsis
- * The paper introduces a new method using endoglycosidase-assisted peptide mapping combined with mass spectrometry to analyze these non-canonical glycosylation sites, specifically in monoclonal antibodies (mAbs).
- * A case study is presented where this workflow successfully identified a unique glycosite in a mAb, enhancing the efficiency of peptide analysis and enabling better understanding of glycosylation patterns important for therapeutic development.
Proteomic interrogation of complex biomedical samples using the rapid denaturing organic digestion (DOD) method.
J Proteomics
February 2025
Nottingham Trent University, Clifton Lane, Nottingham NG11 8NS, United Kingdom; Vibrat-Ion Ltd, Aberystwyth, Wales, SY23 3AF, United Kingdom.
Article Synopsis
- Current aqueous-based tryptic digestion methods face challenges like long digestion times and variability in peptide identification and sequence coverage.
- The Denaturing Organic Digestion (DOD) method offers a faster alternative, requiring only common solvents and reagents, without needing expensive equipment.
- Results showed that the DOD method generated similar peptide profiles to traditional methods but identified more hydrophilic peptides; longer digestion times improved precision and peptide identification.
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