title: High-throughput analysis of drug dissociation from serum proteins using affinity silica monoliths.
abstract: A noncompetitive peak decay method was used with 1 mm×4.6 mm id silica monoliths to measure the dissociation rate constants (kd) for various drugs with human serum albumin (HSA) and α1-acid glycoprotein (AGP). Flow rates up to 9 mL/min were used in these experiments, resulting in analysis times of only 20-30 s. Using a silica monolith containing immobilized HSA, dissociation rate constants were measured for amitriptyline, carboplatin, cisplatin, chloramphenicol, nortriptyline, quinidine, and verapamil, giving values that ranged from 0.37 to 0.78 s(-1). Similar work with an immobilized AGP silica monolith gave kd values for amitriptyline, nortriptyline, and lidocaine of 0.39-0.73 s(-1). These kd values showed good agreement with values determined for drugs with similar structures and/or affinities for HSA or AGP. It was found that a kd of up to roughly 0.80 s(-1) could be measured by this approach. This information made it possible to obtain a better understanding of the advantages and possible limitations of the noncompetitive peak decay method and in the use of affinity silica monoliths for the high-throughput analysis of drug-protein dissociation.
authors.affiliation: Department of Chemistry, University of Nebraska-Lincoln, Lincoln, NE, USA.
authors.forename: Michelle J
authors.initials: MJ
authors.lastname: Yoo
authors.forename: David S
authors.initials: DS
authors.lastname: Hage
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007700:Kinetics; D009961:Orosomucoid / Q000737:chemistry*; D004364:Pharmaceutical Preparations / Q000097:blood / Q000737:chemistry*; D012709:Serum Albumin / Q000737:chemistry*; D012822:Silicon Dioxide / Q000737:chemistry*
chemical_list: D009961:Orosomucoid; D004364:Pharmaceutical Preparations; D012709:Serum Albumin; D012822:Silicon Dioxide