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

Kyla F Ortved; Larry Alward; Bobby Cowles; Renata Linardi; Dhvani Barot; Alex Usimaki; Joseph R Fedie; Deb Amodie; Laurie R Goodrich

doi:10.3389/fvets.2024.1335972

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 Feb 9:11:1335972. doi: 10.3389/fvets.2024.1335972. eCollection 2024.

Authors

Kyla F Ortved¹, Larry Alward², Bobby Cowles³, Renata Linardi¹, Dhvani Barot¹, Alex Usimaki¹, Joseph R Fedie², Deb Amodie⁴, Laurie R Goodrich⁵

Affiliations

¹ New Bolton Center, University of Pennsylvania, Kennett, PA, United States.
² Veterinary Medicine Research and Development, Zoetis, Kalamazoo, MI, United States.
³ Equine Technical Services, Zoetis, Parsippany, NJ, United States.
⁴ Outcomes Research, Zoetis, Parsippany, NJ, United States.
⁵ Orthopaedic Research Center, Translational Medicine Institute, College of Veterinary Medicine and Biomedical Science, Colorado State University, Ft Collins, CO, United States.

Abstract

Introduction: Alpha 2 macroglobulin (A2M), a multi-functional protein in the plasma protease inhibitor class, regulates proinflammatory cytokines and the clearance of chondrodestructive enzymes in cases of joint injury and osteoarthritis (OA). The purpose of this study was to compare A2M concentrations in equine plasma samples processed by three commercial devices developed for stall-side regenerative joint therapy.

Methods: Plasma samples were obtained from healthy adult horses (N = 13). Mass spectrometry analysis was used to determine the concentration of protein analytes in each sample. Selected reaction monitoring measured a specific A2M peptide as a surrogate of the whole A2M protein. A2M concentrations produced by each test device were compared for two sample types: a pre-concentrate or platelet-poor (PP) component and a final component for use in the horse.

Results: There was no significant difference (p > 0.05) in the geometric mean (GM) concentration of A2M in the final concentration samples produced by the Alpha2EQ^® device (N horses = 13) and the single-centrifugation PP samples produced by the Pro-Stride^® APS (autologous protein solution) device (N = 13) and the Restigen^® PRP (platelet-rich plasma) device (N = 11). When A2M content in final concentration samples produced by each device was compared, the Pro-Stride APS and Restigen PRP samples had significantly greater GM A2M content (p < 0.0001) compared to the Alpha2EQ samples, and the Pro-Stride APS final concentration samples had significantly greater GM A2M concentration (p < 0.0001) versus that for the Restigen PRP final samples.

Discussion: This comparison demonstrated that the volume and A2M concentration of an Alpha2EQ final concentrate are no different than the volume and concentration of A2M in the PP from Pro-Stride or Restigen devices.

Keywords: alpha 2 macroglobulin; equine; orthobiologics; protease inhibitor; proteomic; regenerative joint disease therapy; selected reaction monitoring.

Grants and funding

The author(s) declare financial support was received for the research, authorship, and/or publication of this article. This study received funding from Zoetis LCC. The funder was involved in the study design, analysis, interpretation of data, the writing of this article, or the decision to submit it for publication. The funder was not involved in sample collection.