title: Unravelling nonspecific adsorption of complex protein mixture on surfaces with SPR and MS.
abstract: Characterization of protein adsorption to surfaces has implications from biosensing to protective biocoatings. While research studies have principally focused on determining the magnitude of protein adsorption to surfaces, the proteins involved in the process remains only broadly identified and has not been investigated on several surfaces. To further elucidate the nonspecific adsorption process of serum to surfaces, surface plasmon resonance (SPR) and matrix assisted laser desorption ionization mass spectrometry (MALDI-MS) were used in combination to obtain quantitative and qualitative information about the process of protein adsorption to surfaces. To validate the technique, crude serum was nonspecifically adsorbed on four self-assembled monolayer (SAM) on gold: 16-mercaptohexadecanoic acid (16-MHA), 11-mercaptoundecane(ethylene glycol)3-COOH (PEG), 3-MPA-LHDLHD-OH, and 3-MPA-HHHDD-OH. Direct MS analysis of the nonspecifically adsorbed proteins suggested the presence of a variety of protein (BSA, IgG, and apolipoprotein A-1). Performing a trypsin digestion of the nonspecifically adsorbed proteins confirmed the presence of BSA and apolipoprotein A-1 and further revealed the complexity of the process by detecting the presence of complement C3, SHC-transforming protein 1, and kininogen 2. The level of nonspecific adsorption on different surfaces measured by SPR sensing directly correlated with the intensity of the serum protein and indirectly with the tryptic peptides measured by MS. Detailed analysis of the BSA peptides digested on 16-MHA and for BSA digested in solution was used to investigate the orientation of BSA on this surface. The combination of SPR and MS allows the quantitative and qualitative understanding of protein adsorption processes to surfaces.
authors.affiliation: Département de Chimie, Université de Montréal , C.P. 6128 Succursale Centre-Ville, Montreal, Quebec Canada , H3C 3J7.
authors.forename: Julien
authors.initials: J
authors.lastname: Breault-Turcot
authors.forename: Pierre
authors.initials: P
authors.lastname: Chaurand
authors.forename: Jean-Francois
authors.initials: JF
authors.lastname: Masson
mesh_terms: D000327:Adsorption; D045424:Complex Mixtures; D013058:Mass Spectrometry / Q000379:methods*; D011506:Proteins / Q000737:chemistry*; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D045424:Complex Mixtures; D011506:Proteins