title: Modified peptide monolayer binding His-tagged biomolecules for small ligand screening with SPR biosensors.
abstract: A peptide self-assembled monolayer (SAM) was designed to bind His-tagged biomolecules for surface plasmon resonance (SPR) bioanalysis, which was applied for the determination of K(d) for small ligand screening against CD36. Nonspecific adsorption could be minimized using penta- and hexa-peptide monolayers. In particular, monolayers consisting of 3-mercaptopropionyl-leucinyl-histidinyl-aspartyl-leucinyl-histidinyl-aspartic acid (3-Mpa-LHDLHD) exhibited little (12 ng cm(-2)) nonspecific adsorption in crude serum. Modification of this peptide monolayer with Nα,Nα-bis(carboxymethyl)-L-lysine gave a surface competent for binding His-tagged proteins, as demonstrated using enzyme (human dihydrofolate reductase), protein/antibody and receptor (CD36) examples. Immobilization featured chelation of copper and the His-tagged protein by the peptide monolayer, which could be recycled by removing the copper using imidazole washes prior to reuse.
authors.affiliation: Département de Chimie, Université de Montréal, C. P. 6128 Succ. Centre-Ville, Montréal, Qc, Canada H3C 3J7.
authors.forename: Olivier R
authors.initials: OR
authors.lastname: Bolduc
authors.forename: Patrick
authors.initials: P
authors.lastname: Lambert-Lanteigne
authors.forename: Damien Y
authors.initials: DY
authors.lastname: Colin
authors.forename: Sandy Shuo
authors.initials: SS
authors.lastname: Zhao
authors.forename: Caroline
authors.initials: C
authors.lastname: Proulx
authors.forename: Damien
authors.initials: D
authors.lastname: Boeglin
authors.forename: William D
authors.initials: WD
authors.lastname: Lubell
authors.forename: Joelle N
authors.initials: JN
authors.lastname: Pelletier
authors.forename: James
authors.initials: J
authors.lastname: Féthière
authors.forename: Huy
authors.initials: H
authors.lastname: Ong
authors.forename: Jean-Francois
authors.initials: JF
authors.lastname: Masson
mesh_terms: D000327:Adsorption; D000595:Amino Acid Sequence; D000818:Animals; D018955:CD36 Antigens / Q000378:metabolism*; D002417:Cattle; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006639:Histidine / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D008024:Ligands; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D054852:Small Molecule Libraries / Q000494:pharmacology*; D020349:Surface Plasmon Resonance / Q000379:methods*; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry / Q000378:metabolism
chemical_list: D018955:CD36 Antigens; D055767:Immobilized Proteins; D008024:Ligands; D010455:Peptides; D054852:Small Molecule Libraries; D006639:Histidine; D013762:Tetrahydrofolate Dehydrogenase