Modified peptide monolayer binding His-tagged biomolecules for small ligand screening with SPR biosensors |
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Authors: | Bolduc Olivier R Lambert-Lanteigne Patrick Colin Damien Y Zhao Sandy Shuo Proulx Caroline Boeglin Damien Lubell William D Pelletier Joelle N Féthière James Ong Huy Masson Jean-Francois |
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Affiliation: | Département de Chimie, Université de Montréal, C. P. 6128 Succ. Centre-Ville, Montréal, Qc, Canada H3C 3J7. |
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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. |
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