A Pilot Proteomic Analysis of Tear Fluid in Domestic Cats with and Without Conjunctivitis Using MALDI–TOF/TOF Mass Spectrometry

Feline conjunctivitis is a common ocular disorder; however, the molecular composition of feline tear fluid and its alterations during ocular surface inflammation remain poorly characterized. This pilot study aimed to explore the tear proteome of cats with conjunctivitis using matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI–TOF/TOF MS) and to compare findings with those from clinically healthy cats. Tear samples were collected using Schirmer tear test strips from healthy cats and cats diagnosed with conjunctivitis. Total protein concentration was measured by ultraviolet absorbance spectrophotometry, and tear proteins were separated by SDS–polyacrylamide gel electrophoresis, followed by in-gel trypsin digestion and MALDI–TOF/TOF MS analysis. Nine distinct tear proteins were identified, including antimicrobial and immune-related components such as lactoperoxidase, lactotransferrin, albumin, and immunoglobulin A constant region. Lactoperoxidase and SBP1 were identified in feline tear fluid for the first time. No proteins uniquely associated with conjunctivitis were detected. The mean total tear protein concentration was numerically higher in cats with conjunctivitis (13.06 ± 0.75 mg/mL) than in healthy cats (9.69 ± 0.67 mg/mL); however, this difference did not reach statistical significance (p = 0.095) and should be interpreted cautiously given the limited sample size. This pilot study provides preliminary insights into tear protein profiles in cats with conjunctivitis and highlights the need for larger quantitative investigations. These findings provide a preliminary framework for future studies aimed at further characterizing molecular alterations associated with feline ocular surface disorders.