Serum concentrations of canine interleukin-1 receptor antagonist protein in healthy dogs after incubation using an autologous serum processing system

• Published in: Research in veterinary science Vol. 101; pp. 28 - 33
• Main Authors: Huggins, S.S., Suchodolski, J.S., Bearden, R.N., Steiner, J.M., Saunders, W.B.
• Format: Journal Article
• Language: English
• Published: England Elsevier Ltd 01.08.2015; Elsevier Limited
• Subjects: Animals; Arthritis; Autologous serum; Blood Chemical Analysis - methods; Blood Chemical Analysis - veterinary; Canine interleukin-1 receptor antagonist; Clinical trials; Cytokines; Dogs - blood; ELISA; Enzyme-Linked Immunosorbent Assay - methods; Enzyme-Linked Immunosorbent Assay - veterinary; Gene expression; Horses; Interleukin 1 Receptor Antagonist Protein - blood; Orthobiologics; Osteoarthritis; Proteins; Receptors, Interleukin-1 - antagonists & inhibitors; Serum - chemistry; Studies; Veterinary medicine; Canine interleukin-1 receptor antagonist; Autologous serum; Osteoarthritis; Orthobiologics; ELISA
• ISSN: 0034-5288; 1532-2661
• DOI: 10.1016/j.rvsc.2015.05.012

The objectives of this study were to optimize and validate a canine IL-1RA ELISA using commercially available reagents and to determine the effect of an autologous serum processing system (IRAP II) on IL-1RA concentrations in canine serum. The clinical detection limit of the optimized ELISA was 188.8 to 39,965.6pg/mL. The observed-to-expected ratio (O:E) for three serial dilutions for four serum samples ranged from 109.6 to 132.2%. The O:E for four serum samples spiked with four concentrations of canine IL-1 RA ranged from 98.7 to 114.3%. Coefficients of variances for intra- and interassay variability ranged from 1.4 to 3.0 and 6.3 to 9.8, respectively. The ELISA was sensitive, linear, accurate, precise, and reproducible. Mean±SD serum concentration of IL-1RA in 12 healthy dogs was 396.6±208.0pg/mL. There was a significant increase in IL-1RA when blood was incubated in the IRAP II system (15,955.0±6421.0pg/mL, P<0.0001). •Autologous serum systems that enrich IL-1RA may prove useful for treating canine osteoarthritis.•A linear, accurate, precise, and repeatable canine IL-1RA ELISA has yet to be described.•We report the optimization and validation of a canine IL-1RA ELISA for use on canine serum.•Using this ELISA, IL-1 RA levels in healthy canine blood are 396.6±208.0pg/mL (mean±SD).•IL-1 levels increased to 15,955.0±6421.0pg/mL after treatment with an autologous serum system.