Modified peptide monolayer binding His-tagged biomolecules for small ligand screening with SPR biosensors

• Published in: Analyst (London) Vol. 136; no. 15; pp. 3142 - 3148
• Main Authors: BOLDUC, Olivier R, LAMBERT-LANTEIGNE, Patrick, MASSON, Jean-Francois, COLIN, Damien Y, SHUO ZHAO, Sandy, PROULX, Caroline, BOEGLIN, Damien, LUBELL, William D, PELLETIER, Joelle N, FETHIERE, James, ONG, Huy
• Format: Journal Article
• Language: English
• Published: Cambridge Royal Society of Chemistry 07.08.2011
• Subjects: Adsorption; Amino Acid Sequence; Analytical chemistry; Animals; Biological and medical sciences; Biosensors; Biotechnology; Cattle; CD36 Antigens - metabolism; Chemistry; Drug Evaluation, Preclinical - methods; Exact sciences and technology; Fundamental and applied biological sciences. Psychology; General, instrumentation; Histidine - chemistry; Histidine - metabolism; Humans; Immobilized Proteins - chemistry; Immobilized Proteins - metabolism; Ligands; Methods. Procedures. Technologies; Peptides - chemistry; Peptides - metabolism; Protein Binding; Small Molecule Libraries - pharmacology; Surface Plasmon Resonance - methods; Tetrahydrofolate Dehydrogenase - chemistry; Tetrahydrofolate Dehydrogenase - metabolism; Various methods and equipments; Self-assembled layer; Human; Monolayer; Peptides; Antibody; Enzyme; Ligand; Immobilization; Chemical sensor; Chemical enrichment; Design; Screening; Binding protein; Adsorption; Lysine; Biosensor; Imidazole; Chelation; Serum; Aspartic acid; Surface plasmon resonance; Copper
• ISSN: 0003-2654; 1364-5528
• DOI: 10.1039/c1an15235a

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.