Proteomic Analysis of Tears and Conjunctival Cells Collected with Schirmer Strips Using timsTOF Pro: Preanalytical Considerations
Murat Akkurt Arslan,
Ioannis Kolman,
Cédric Pionneau,
Solenne Chardonnet,
Romain Magny,
Christophe Baudouin,
Françoise Brignole-Baudouin,
Karima Kessal
Affiliations
Murat Akkurt Arslan
Institut National de la Santé et de la Recherche Médicale INSERM UMR 968, CNRS UMR 7210, Institut de la Vision, IHU ForeSight, Sorbonne Université UM80, 75012 Paris, France
Ioannis Kolman
Institut National de la Santé et de la Recherche Médicale INSERM UMR 968, CNRS UMR 7210, Institut de la Vision, IHU ForeSight, Sorbonne Université UM80, 75012 Paris, France
Cédric Pionneau
Institut National de la Santé et de la Recherche Médicale INSERM, UMS PASS, Plateforme Post-Génomique de la Pitié Salpêtrière (P3S), Sorbonne Université, 75013 Paris, France
Solenne Chardonnet
Institut National de la Santé et de la Recherche Médicale INSERM, UMS PASS, Plateforme Post-Génomique de la Pitié Salpêtrière (P3S), Sorbonne Université, 75013 Paris, France
Romain Magny
Institut National de la Santé et de la Recherche Médicale INSERM UMR 968, CNRS UMR 7210, Institut de la Vision, IHU ForeSight, Sorbonne Université UM80, 75012 Paris, France
Christophe Baudouin
Institut National de la Santé et de la Recherche Médicale INSERM UMR 968, CNRS UMR 7210, Institut de la Vision, IHU ForeSight, Sorbonne Université UM80, 75012 Paris, France
Françoise Brignole-Baudouin
Institut National de la Santé et de la Recherche Médicale INSERM UMR 968, CNRS UMR 7210, Institut de la Vision, IHU ForeSight, Sorbonne Université UM80, 75012 Paris, France
Karima Kessal
Institut National de la Santé et de la Recherche Médicale INSERM UMR 968, CNRS UMR 7210, Institut de la Vision, IHU ForeSight, Sorbonne Université UM80, 75012 Paris, France
This study aimed to investigate the human proteome profile of samples collected from whole (W) Schirmer strips (ScS) and their two parts—the bulb (B) and the rest of the strip (R)—with a comprehensive proteomic approach using a trapped ion mobility mass spectrometer, the timsTOF Pro. Eight ScS were collected from two healthy subjects at four different visits to be separated into three batches, i.e., 4W, 4B, and 4R. In total, 1582 proteins were identified in the W, B, and R batches. Among all identified proteins, binding proteins (43.4%) and those with catalytic activity (42.2%) constituted more than 80% of the molecular functions. The most represented biological processes were cellular processes (31.2%), metabolic processes (20.8%), and biological regulation (13.1%). Enzymes were the most represented protein class (41%), consisting mainly of hydrolases (47.5%), oxidoreductases (22.1%), and transferases (16.7%). The bulb (B), which is in contact with the conjunctiva, might collect both tear and cell proteins and therefore promote the identification of more proteins. Processing B and R separately before mass spectrometry (MS) analysis, combined with the high data acquisition speed and the addition of ion-mobility-based separation in the timsTOF Pro, can bring a new dimension to biomarker investigations of a limited sample such as tear fluid.
