Severe equine asthma (SEA) is a common, chronic respiratory disease of horses characterized by hyperreactivity to hay dust which has many similarities to severe neutrophilic asthma in humans. A 2D immunoblot approach was used to identify quantitative differences in the recognition of the two-dimensional gel electrophoresis (2-DE) pattern between asthmatic and helthy horses´sera. Differentially regulated spots were cut from a 2-D gel and used for in-gel digestion followed by LC-MS to identify possible antigens.

Peptides obtained after tryptic in-gel digest or FASP were analyzed with a nanoACQUITY Ultra Performance LC™ coupled to a Q-TOF SYNAPT G2-Si instrument. For measurments a high-definition data-dependent acquisition approach (HD-DDA) for top 6 ions was used. Followed by data analysis with Mascot Distiller (Version 2.8.1.0) and Mascot search engine (Version 2.7.0).

103 tryptic in-gel digested samples, 1 tryptic filter-aided sample preparation sample (100 ng on column)