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    Proteomic Analysis of Biomaterial Surfaces after Contacting with Body Fluids by MALDI-ToF Mass Spectroscopy

    Year: 2020

    Journal: Coatings, Volume 10, JAN

    Authors: Hirohara, Makoto; Maekawa, Tatsuhiro; Mondarte, Evan Angelo Quimada; Nyu, Takashi; Mizushita, Yoshiki; Hayashi, Tomohiro

    Keywords: protein identification; matrix-assisted laser desorption/ionization-time of flight; proteomics; quartz crystal microbalance; self-assembled monolayer

    We developed a method to identify proteins adsorbed on solid surfaces from a solution containing a complex mixture of proteins by using Matrix-Assisted Laser Desorption/Ionization-Time of Flight mass (MALDI-ToF mass) spectroscopy. In the method, we performed all procedures of peptide mass fingerprint method including denaturation, reduction, alkylation, digestion, and spotting of matrix on substrates. The method enabled us to avoid artifacts of pipetting that could induce changes in the composition. We also developed an algorithm to identify the adsorbed proteins. In this work, we demonstrate the identification of proteins adsorbed on self-assembled monolayers (SAMs). Our results show that the composition of proteins on the SAMs critically depends on the terminal groups of the molecules constituting the SAMs, indicating that the competitive adsorption of protein molecules is largely affected by protein-surface interaction. The method introduced here can provide vital information to clarify the mechanism underlying the responses of cells and tissues to biomaterials.
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