title: A Surface Plasmon Resonance Spectroscopy Method for Characterizing Small-Molecule Binding to Nerve Growth Factor.
abstract: Small-molecule inhibitors have been previously investigated to identify possible therapeutics for the treatment of chronic pain. In the present study, known nerve growth factor (NGF) inhibitors identified by (125)I-NGF binding were characterized using affinity and binding evaluations by surface plasmon resonance (SPR) spectroscopy. A novel strategy for characterizing NGF inhibitors was used to determine the binding affinity (KD) and saturation ability of each compound with immobilized NGF. Seventy-four percent of compounds screened demonstrated a positive binding event to NGF. A KD less than 10 μM and a percent saturation greater than 50% were used as thresholds to identify inhibitors that would warrant further investigation. This study details for the first time a methodology that can be used to directly characterize the binding event between small-molecule inhibitors and NGF.
authors.affiliation: Laurentian University, Biomolecular Sciences Program, Sudbury, ON, Canada.
authors.forename: Allison E
authors.initials: AE
authors.lastname: Kennedy
authors.affiliation: Laurentian University, Biology Department, Sudbury, ON, Canada.
authors.forename: Kristen S
authors.initials: KS
authors.lastname: Sheffield
authors.affiliation: Northern Ontario School of Medicine, Sudbury, ON, Canada.
authors.forename: Joseph K
authors.initials: JK
authors.lastname: Eibl
authors.affiliation: GE Healthcare Life Sciences, Piscataway, NJ, USA.
authors.forename: Michael B
authors.initials: MB
authors.lastname: Murphy
authors.affiliation: Sussex Research, Ottawa, ON, Canada.
authors.forename: Rahul
authors.initials: R
authors.lastname: Vohra
authors.affiliation: Laurentian University, Biomolecular Sciences Program, Sudbury, ON, Canada.
authors.forename: John A
authors.initials: JA
authors.lastname: Scott
authors.affiliation: Laurentian University, Biology Department, Sudbury, ON, Canada Northern Ontario School of Medicine, Sudbury, ON, Canada gross@nosm.ca.
authors.forename: Gregory M
authors.initials: GM
authors.lastname: Ross
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D007700:Kinetics; D020932:Nerve Growth Factor / Q000037:antagonists & inhibitors*; D011485:Protein Binding / Q000502:physiology*; D051381:Rats; D054852:Small Molecule Libraries / Q000494:pharmacology*; D013057:Spectrum Analysis / Q000379:methods; D020349:Surface Plasmon Resonance / Q000379:methods
chemical_list: D054852:Small Molecule Libraries; D020932:Nerve Growth Factor
keywords: NGF; SPR; biosensor technology; neurotrophin; pain therapeutics