title: Diffusion-driven device for a high-resolution dose-response profiling of combination chemotherapy.
abstract: Combination therapies have proven vital in the fight against HIV and cancer. However, the identification and optimization of such combination therapies is largely experience driven and an activity of clinicians rather than of systematic screening efforts. Here we present a diffusion device, compatible with the format of a 12-well microtiter plate, to create and test all possible mixtures of two substances with only two pipetting steps. Applications to the testing of different drug combinations and the parallel screening of different leukemia cell lines as well as primary patient cells are presented. The diffusion device yields qualitatively and quantitatively comparable results to an MTT viability assay conducted in a standard 96-well format albeit with a tremendous reduction of processing steps. In addition, a fluorescence-based annexin V binding assay of cell death was implemented. Next to the reduction of processing steps, the diffusion device constitutes a considerable assay miniaturization that overcomes the problems typically associated with miniaturization as a consequence of small sample volumes. Given its ease of handling, the device will greatly advance the development and optimization of combination drugs and the identification of optimum drug combinations in personalized medicine.
authors.affiliation: Department of Biochemistry, Nijmegen Centre for Molecular Life Sciences, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
authors.forename: Alexander
authors.initials: A
authors.lastname: Ganser
authors.forename: Günter
authors.initials: G
authors.lastname: Roth
authors.forename: Joost C
authors.initials: JC
authors.lastname: van Galen
authors.forename: Janneke
authors.initials: J
authors.lastname: Hilderink
authors.forename: Joost J G
authors.initials: JJ
authors.lastname: Wammes
authors.forename: Ingo
authors.initials: I
authors.lastname: Müller
authors.forename: Frank N
authors.initials: FN
authors.lastname: van Leeuwen
authors.forename: Karl-Heinz
authors.initials: KH
authors.lastname: Wiesmüller
authors.forename: Roland
authors.initials: R
authors.lastname: Brock
mesh_terms: D000227:Adenine Nucleotides / Q000494:pharmacology; D017304:Annexin A5 / Q000737:chemistry / Q000494:pharmacology; D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D001087:Arabinonucleosides / Q000494:pharmacology; D016923:Cell Death / Q000187:drug effects; D000077866:Clofarabine; D004058:Diffusion; D004305:Dose-Response Relationship, Drug; D004317:Doxorubicin / Q000494:pharmacology; D004359:Drug Therapy, Combination; D005456:Fluorescent Dyes / Q000737:chemistry; D006801:Humans; D008904:Miniaturization; D010718:Phosphatidylserines / Q000494:pharmacology; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000188:drug therapy; D018675:Toxicity Tests / Q000295:instrumentation*; D014407:Tumor Cells, Cultured
chemical_list: D000227:Adenine Nucleotides; D017304:Annexin A5; D001087:Arabinonucleosides; D005456:Fluorescent Dyes; D010718:Phosphatidylserines; D000077866:Clofarabine; D004317:Doxorubicin