Multiplexed immunoassay for a serum autoantibody biomarker panel in diagnostic and prognostic prediction of canine mammary tumors

• Published in: The Veterinary quarterly Vol. 45; no. 1; pp. 1 - 12
• Main Authors: Wu, Chih-Ching, Chang, Chia-Yu, Chou, Pei-Yi, Chan, Xiu-Ya, Huang, Chun-Chueh, Yang, Youngsen, Liu, Hao-Ping
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01.12.2025; Taylor & Francis Group
• Subjects: Animals; Autoantibodies - blood; autoantibody; biomarker; Biomarkers, Tumor - blood; Canine mammary tumor; Dog Diseases - blood; Dog Diseases - diagnosis; Dog Diseases - immunology; Dogs; Female; Immunoassay - methods; Immunoassay - veterinary; Mammary Neoplasms, Animal - blood; Mammary Neoplasms, Animal - diagnosis; Mammary Neoplasms, Animal - immunology; multiplexed assay; Prognosis; Sensitivity and Specificity; serum; autoantibody; multiplexed assay; biomarker; serum; Canine mammary tumor
• ISSN: 0165-2176; 1875-5941; 1875-5941
• DOI: 10.1080/01652176.2024.2435978

Canine mammary tumor (CMT) is a prevalent and destructive disease often diagnosed at an advanced stage, leading to poor outcomes. Currently, there is a lack of effective biomarkers for early detection and prognostic prediction of CMT. To improve CMT detection, we established a multiplexed immunoassay using a fluorescence bead-based suspension array system to measure serum levels of autoantibodies against four CMT-associated proteins (AGR2, HAPLN1, IGFBP5, and TYMS) in CMT patients. Our data revealed that serum levels of the four autoantibodies (anti-AGR2, anti-HAPLN1, anti-IGFBP5, and anti-TYMS) were significantly elevated in CMT patients (  = 158) compared to healthy individuals (  = 39). Notably, serum levels of anti-AGR2, anti-HAPLN1, and anti-TYMS in the dogs with stage I CMT (  = 56) were higher than those in the healthy group. Using a marker panel consisting of the four autoantibodies for detecting malignant CMT (  = 125) achieved a sensitivity of 50.4% and a specificity of 90%. Furthermore, higher levels of anti-AGR2, anti-HAPLN1, anti-IGFBP5, and anti-TYMS were associated with poorer survival in CMT patients. Collectively, we established a multiplexed immunoassay platform to detect serum autoantibodies and demonstrated that a tailored autoantibody marker panel shows potential clinical applicability for the diagnosis and prognosis of CMT.