Validation of commercial ELISAs for quantifying anabolic growth factors and cytokines in canine ACD-A anticoagulated plasma.
Platelet-rich plasma has been studied extensively in dogs, but validation of enzyme-linked immunosorbent assays (ELISAs) for quantifying anabolic growth factors and inflammatory cytokines in canine plasma prepared with citrate-based anticoagulants is not available. We performed a validation of commercial ELISAs for transforming growth factor-beta 1 (TGF-β1), platelet-derived growth factor-BB (PDGF-BB), vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-1β) for use with canine plasma prepared with acid-citrate-dextrose, solution A (ACD-A). Platelet-poor plasma (PPP) anticoagulated with ACD-A as well as PPP anticoagulated with ACD-A and spiked with the relevant canine recombinant proteins were evaluated with each ELISA to calculate the efficiency of spike recovery. Replicates of the spiked PPP were also assessed in 2 additional assays to quantify intra-assay and interassay precision. The efficiency of spike recovery was within 75-125% of the expected concentration for the TGF-β1, PDGF-BB, and VEGF ELISAs. The intra- and interassay variability were <25% for the TGF-β1, PDGF-BB, VEGF, and TNF-α ELISAs. The TGF-β1, PDGF-BB, and VEGF ELISAs demonstrate acceptable efficiency of spike recovery and intra- and interassay variability, whereas the TNF-α and IL-1β ELISAs did not meet industry standards of performance with ACD-A anticoagulated canine plasma.
Department of Small Animal Medicine and Surgery (Birdwhistell, Basinger, Hayes, Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Regenerative Bioscience Center (Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Large Animal Medicine and Surgery (Norton, Hurley), University of Georgia, College of Veterinary Medicine, Athens, GA.
Kate
K
Birdwhistell
Department of Small Animal Medicine and Surgery (Birdwhistell, Basinger, Hayes, Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Regenerative Bioscience Center (Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Large Animal Medicine and Surgery (Norton, Hurley), University of Georgia, College of Veterinary Medicine, Athens, GA.
Robert
R
Basinger
Department of Small Animal Medicine and Surgery (Birdwhistell, Basinger, Hayes, Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Regenerative Bioscience Center (Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Large Animal Medicine and Surgery (Norton, Hurley), University of Georgia, College of Veterinary Medicine, Athens, GA.
Brian
B
Hayes
Department of Small Animal Medicine and Surgery (Birdwhistell, Basinger, Hayes, Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Regenerative Bioscience Center (Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Large Animal Medicine and Surgery (Norton, Hurley), University of Georgia, College of Veterinary Medicine, Athens, GA.
Natalie
N
Norton
Department of Small Animal Medicine and Surgery (Birdwhistell, Basinger, Hayes, Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Regenerative Bioscience Center (Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Large Animal Medicine and Surgery (Norton, Hurley), University of Georgia, College of Veterinary Medicine, Athens, GA.
David J
DJ
Hurley
Department of Small Animal Medicine and Surgery (Birdwhistell, Basinger, Hayes, Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Regenerative Bioscience Center (Franklin), University of Georgia, College of Veterinary Medicine, Athens, GA.|Large Animal Medicine and Surgery (Norton, Hurley), University of Georgia, College of Veterinary Medicine, Athens, GA.
Samuel P
SP
Franklin
D000818:Animals; D000925:Anticoagulants; D000077214:Becaplermin; D016207:Cytokines / Q000097:blood*; D004285:Dogs; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary*; D036341:Intercellular Signaling Peptides and Proteins / Q000097:blood*; D053657:Platelet-Rich Plasma / Q000737:chemistry*; D020574:Proto-Oncogene Proteins c-sis / Q000097:blood; D015203:Reproducibility of Results; D045165:Testosterone Congeners / Q000097:blood; D053773:Transforming Growth Factor beta1 / Q000097:blood; D014409:Tumor Necrosis Factor-alpha / Q000097:blood; D042461:Vascular Endothelial Growth Factor A / Q000097:blood
D000925:Anticoagulants; D016207:Cytokines; D036341:Intercellular Signaling Peptides and Proteins; D020574:Proto-Oncogene Proteins c-sis; D045165:Testosterone Congeners; D053773:Transforming Growth Factor beta1; D014409:Tumor Necrosis Factor-alpha; D042461:Vascular Endothelial Growth Factor A; D000077214:Becaplermin
Anabolic growth factors; dogs; enzyme-linked immunosorbent assay; inflammatory cytokines; platelet-rich plasma