title: Simple method provides resolution of albumin, lipoprotein, free fraction, and chylomicron to enhance the utility of protein binding assays.
abstract: Medicinal chemists have been encouraged in recent years to embrace high speed protein binding assays. These methods employ dialysis membranes in 96-well format or spin filters. Membrane-based methods do not separate lipoprotein binding from albumin binding and introduce interference despite membrane binding controls. Ultracentrifugation methods, in contrast, do not introduce interference if density gradients can be avoided and they resolve lipoprotein from albumin. A new generation of compact, fast ultracentrifuges facilitates the rapid and fully informative separation of plasma into albumin, albumin/fatty acid complex, lipoprotein, protein-free, and chylomicron fractions with no need of salt or sugar density gradients. We present a simple and fast ultracentrifuge method here for two platinum compounds and a taxane that otherwise bound irreversibly to dialysis membranes and which exhibited distinctive lipoprotein binding behaviors. This new generation of ultracentrifugation methods underscores a need to further discuss protein binding assessments as they relate to medicinal chemistry efforts.
authors.affiliation: Drug Metabolism & Pharmacokinetics, Blend Therapeutics , 134 Coolidge Avenue, Watertown, Massachusetts 02472, United States.
authors.forename: Adam H
authors.initials: AH
authors.lastname: Brockman
authors.forename: Haley R
authors.initials: HR
authors.lastname: Oller
authors.forename: Benoît
authors.initials: B
authors.lastname: Moreau
authors.forename: Kristina
authors.initials: K
authors.lastname: Kriksciukaite
authors.forename: Mark T
authors.initials: MT
authors.lastname: Bilodeau
mesh_terms: D000418:Albumins / Q000737:chemistry*; D001952:Bridged-Ring Compounds / Q000737:chemistry; D002626:Chemistry, Pharmaceutical; D003956:Dialysis; D008074:Lipoproteins / Q000737:chemistry*; D015394:Molecular Structure; D009944:Organoplatinum Compounds / Q000737:chemistry; D011485:Protein Binding; D043823:Taxoids / Q000737:chemistry; D014461:Ultracentrifugation
chemical_list: D000418:Albumins; D001952:Bridged-Ring Compounds; D008074:Lipoproteins; D009944:Organoplatinum Compounds; D043823:Taxoids; C080625:taxane