Livestock, sustainable and herd of cattle on a farm in the countryside for eco friendly environment. Agriculture, animals and cows for meat, dairy or beef trade production industry in a grass field
Veterinary medicine increasingly demands the same analytical rigor applied in human clinical research. Whether researchers are characterizing the protein composition of an equine regenerative therapy, detecting drug residues in livestock or profiling pathogen virulence factors in production animals, the questions are complex, and the tools must be equal to them.
Mass spectrometry (MS) has become the leading platform for quantitative proteomics in animal health research. Its ability to identify and measure proteins with extraordinary specificity, without the species-specific antibody constraints of immunoassays, makes it uniquely suited to the broad demands of veterinary and translational science. In particular, selected reaction monitoring (SRM) mass spectrometry delivers absolute protein quantification in complex biological matrices, opening new doors for veterinary diagnostics and therapeutic characterization across species.
Two application areas illustrate this impact especially well: toxicology diagnostics, where MS enables rapid, multiplexed detection of harmful agents across animal populations, and therapeutic protein characterization, where proteomic profiling is informing next-generation veterinary biologics and regenerative treatment strategies.
Advancing Veterinary Toxicology with Mass Spectrometry
Toxicological surveillance in animal populations protects herd health, preserves food safety and safeguards the integrity of animal-derived products. Traditional immunoassay-based screening is fast but frequently lacks the specificity to distinguish structurally related toxins, metabolites or residues at trace levels. Mass spectrometry closes that gap.
LC-MS/MS methods are now widely adopted as the preferred regulatory approach for residue analysis across veterinary matrices, including mycotoxins in feed, antibiotic residues in meat and milk, antiparasitic agents in aquaculture systems and environmental contaminants across livestock species (1). The ability to build multiplexed panels that simultaneously quantify dozens of analytes from a single extraction is transforming throughput and turnaround in veterinary diagnostic laboratories.
Meanwhile, emerging proteomics-based toxicology approaches are adding another dimension. Protein biomarkers of organ stress, including markers of hepatotoxicity, nephrotoxicity and immune dysregulation, can in principle be quantified directly from serum or urine by SRM-MS without species-specific antibody reagents. This capability is especially promising for companion animals, aquaculture species and exotic animals where validated immunoassay platforms are scarce.
Across the field, applications span early drug safety screening in target animal species, environmental exposure monitoring in livestock and wildlife, and residue confirmation testing that holds up to regulatory review.
Featured Study: Comparing Equine Orthobiologic Devices with SRM-MS
A 2024 study by Ortved et al., published in Frontiers in Veterinary Science, offers a compelling example of what MS-based proteomics can do for veterinary therapeutic development (2). The researchers used SRM mass spectrometry alongside ELISA to compare alpha-2-macroglobulin (A2M) concentrations across three commercial stall-side regenerative joint therapy devices: Alpha2EQ (Astaria), Pro-Stride APS (Zoetis) and Restigen PRP (Zoetis).
A2M is a multi-functional protease inhibitor that suppresses proinflammatory cytokines and clears chondrodestructive enzymes, making it a critical biomarker in equine osteoarthritis therapy. SRM mass spectrometry provided a species-independent quantification route, circumventing the limitations of equine-specific ELISA reagents.
The findings show how device design and measurement method interact. By SRM-MS, the A2M concentration in the Alpha2EQ final concentrate was statistically indistinguishable from the platelet-poor plasma fractions of the other two devices (p > 0.05). A head-to-head comparison of the three final products did show significant concentration differences (p < 0.0001). At the same time, the total A2M recovered across all collected fractions did not differ significantly among the devices, and an orthogonal ELISA comparison of the final products likewise found no significant difference. The picture that emerges is less about ranking products and more about how processing steps and the analytical method shape what gets measured (2).
What stands out here is the role of mass spectrometry itself. The study highlights how SRM-MS can provide precise, species-independent protein quantification that enables direct, apples-to-apples comparison of orthobiologic devices. That kind of quantitative clarity is exactly what the field needs to advance product standardization and inform evidence-based treatment decisions in equine practice.
Mass Spectrometry in Veterinary Therapeutic Development
Beyond regenerative medicine, MS-based proteomics is gaining traction in veterinary vaccine and therapeutic research more broadly. Effective veterinary therapeutics, from biologics and vaccines to antimicrobials, require a detailed understanding of the proteins driving disease and recovery. Mass spectrometry enables researchers to characterize pathogen virulence proteins, monitor host immune responses and validate therapeutic mechanisms with a level of precision that antibody-based tools cannot match across diverse animal species.
Streptococcus agalactiae (Group B Streptococcus, GBS) is a good example. In dairy cattle, GBS is a leading cause of contagious mastitis, driving significant production losses (3). Developing targeted veterinary vaccines requires detailed characterization of GBS surface proteins and their interactions with bovine immune effectors. MS-based proteomics is well positioned to advance this work, and several research directions are emerging.
LC-MS/MS-based shaving proteomics, for instance, can identify surface-exposed proteins across bacterial strains and isolates, revealing conserved vaccine antigen candidates. Quantitative proteomics is also being used to characterize host immune proteins recruited during infection, identifying protective mechanisms and immune evasion strategies that inform therapeutic design. And SRM assays targeting pathogen or host proteins offer the potential to monitor therapeutic response and disease progression in veterinary research and emerging clinical applications.
Across all these applications, the analytical consistency and species-agnostic nature of mass spectrometry makes it a preferred platform for rigorous veterinary research.
Supporting the Workflow: Promega Tools for Veterinary Proteomics
From sample preparation through final quantification, the proteomics workflow depends on reagent quality. Promega’s portfolio spans the complete workflow and is designed to perform reliably across the diverse and challenging matrices encountered in veterinary science.
Trypsin Gold, Mass Spectrometry Grade provides the consistent, reproducible peptide digestion that bottom-up proteomics demands, including in complex veterinary matrices like serum, synovial fluid and milk. For proteins that resist standard trypsin digestion, rLys-C and rAsp-N proteases enable multi-enzyme digestion strategies essential for comprehensive sequence coverage of large proteins like A2M.
Stable isotope-labeled peptide standards enable absolute protein quantification by SRM/MRM, the analytical approach used in studies like the Ortved et al. comparison of orthobiologic devices. Maxwell® automated sample prep reduces variability and increases throughput across multi-site studies. For researchers working upstream of proteomics, NanoLuc® bioluminescent reporter assays bridge the gap between discovery proteomics and mechanism-of-action studies in veterinary therapeutic development.
Looking Ahead
The Ortved et al. study is a microcosm of a broader transformation. When researchers needed to compare protein concentrations across commercial equine joint therapy devices, they reached for mass spectrometry because SRM-MS offered species-independent absolute quantification with a defined analytical response that holds up across publication and regulatory review.
The same logic applies across animal health science: mycotoxin residues in a commercial flock, the surface proteome of a mastitis-causing pathogen, the therapeutic protein content of a regenerative blood product. Mass spectrometry provides a single analytical language that works across biological complexity, species and regulatory jurisdictions. As the veterinary field continues to adopt the analytical standards already established in human clinical research, the tools and reagents that support those workflows will only become more important.
Explore Promega’s mass spectrometry solutions for animal health research: Browse the MS Portfolio at promega.com
References
1. European Commission. Commission Implementing Regulation (EU) 2021/808 of 22 March 2021 on the performance of analytical methods for residues of pharmacologically active substances used in food-producing animals and on the interpretation of results. Official Journal of the European Union. L 180, 21 May 2021, p. 84.
2. Ortved KF, Alward L, Cowles B, Linardi R, Barot D, Usimaki A, Fedie JR, Amodie D, Goodrich LR. Use of quantitative mass spectrometry-based proteomics and ELISA to compare the alpha 2 macroglobulin concentration in equine blood-based products processed by three different orthobiologic devices. Front. Vet. Sci. 2024;11:1335972. doi: 10.3389/fvets.2024.1335972
3. Keefe GP. Streptococcus agalactiae mastitis: a review. Can. Vet. J. 1997;38(7):429-437.
Sara Millevolte
