title: Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug-DNA interactions.
abstract: Determination of the DNA-binding reactivity and affinity is an important part of a successful program for the selection of metallodrug candidates. For such assaying, a range of complementary analytical techniques was proposed and tested here using one of few anticancer metal-based drugs that are currently in clinical trials, indazolium trans-[tetrachloridobis(1H-indazole)ruthenate(III), and a DNA oligonucleotide. A high reactivity of the Ru drug was confirmed in affinity capillary electrophoresis (CE) mode, where adduct formation takes place in situ (i.e., in the capillary filled with an oligonucleotide-containing electrolyte). To further characterize the binding kinetics, a drug-oligonucleotide mixture was incubated for a different period of time, followed by ultrafiltration separation into two different in molecular weight fractions (>3 and <3 kDa). The time-dependent distribution profiles of the Ru drug were then assessed by CE-inductively coupled plasma mass spectrometry (ICP-MS), revealing that at least two DNA adducts exist at equilibrium conditions. Using standalone ICP-MS, dominant equilibrium amount of the bound ruthenium was found to occur in a fraction of 5-10 kDa, which includes the oligonucleotide (ca. 6 kDa). Importantly, in all three assays, the drug was used for the first time in in-vitro studies, not in the intact form but as its active species released from the transferrin adduct at simulated cancer cytosolic conditions. This circumstance makes the established analytical platform promising to provide a detailed view on metallodrug targeting, including other possible biomolecules and ex vivo samples.
authors.affiliation: Vernadsky Institute of Geochemistry and Analytical Chemistry, Kosygin St. 19, 119991, Moscow, Russian Federation.
authors.forename: Lidia S
authors.initials: LS
authors.lastname: Foteeva
authors.affiliation: Chair of Analytical Chemistry, Faculty of Chemistry, Warsaw University of Technology, Noakowskiego St. 3, 00-664, Warsaw, Poland. mmatczuk@ch.pw.edu.pl.
authors.forename: Magdalena
authors.initials: M
authors.lastname: Matczuk
authors.affiliation: Chair of Analytical Chemistry, Faculty of Chemistry, Warsaw University of Technology, Noakowskiego St. 3, 00-664, Warsaw, Poland.
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Pawlak
authors.affiliation: Saratov State University, Astrakhanskaya St. 83, 410012, Saratov, Russian Federation.
authors.forename: Svetlana S
authors.initials: SS
authors.lastname: Aleksenko
authors.affiliation: Institute of Microelectronics Technology and High-Purity Materials, Acad. Ossipyan St. 6, 142432, Chernologolovka, Russian Federation.
authors.forename: Sergey V
authors.initials: SV
authors.lastname: Nosenko
authors.affiliation: Institute of Microelectronics Technology and High-Purity Materials, Acad. Ossipyan St. 6, 142432, Chernologolovka, Russian Federation.
authors.forename: Vasily K
authors.initials: VK
authors.lastname: Karandashev
authors.affiliation: Chair of Analytical Chemistry, Faculty of Chemistry, Warsaw University of Technology, Noakowskiego St. 3, 00-664, Warsaw, Poland.
authors.forename: Maciej
authors.initials: M
authors.lastname: Jarosz
authors.affiliation: Vernadsky Institute of Geochemistry and Analytical Chemistry, Kosygin St. 19, 119991, Moscow, Russian Federation.
authors.forename: Andrei R
authors.initials: AR
authors.lastname: Timerbaev
mesh_terms: D004247:DNA / Q000737:chemistry*; D019075:Electrophoresis, Capillary / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D012428:Ruthenium / Q000737:chemistry*
chemical_list: D012428:Ruthenium; D004247:DNA
keywords: Anticancer metallodrugs; Capillary electrophoresis; DNA; ICP-MS