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Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions
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 cur...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Springer Berlin Heidelberg
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352744/ https://www.ncbi.nlm.nih.gov/pubmed/28116493 http://dx.doi.org/10.1007/s00216-017-0186-0 |
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author | Foteeva, Lidia S. Matczuk, Magdalena Pawlak, Katarzyna Aleksenko, Svetlana S. Nosenko, Sergey V. Karandashev, Vasily K. Jarosz, Maciej Timerbaev, Andrei R. |
author_facet | Foteeva, Lidia S. Matczuk, Magdalena Pawlak, Katarzyna Aleksenko, Svetlana S. Nosenko, Sergey V. Karandashev, Vasily K. Jarosz, Maciej Timerbaev, Andrei R. |
author_sort | Foteeva, Lidia S. |
collection | PubMed |
description | 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-017-0186-0) contains supplementary material, which is available to authorized users. |
format | Online Article Text |
id | pubmed-5352744 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Springer Berlin Heidelberg |
record_format | MEDLINE/PubMed |
spelling | pubmed-53527442017-03-27 Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions Foteeva, Lidia S. Matczuk, Magdalena Pawlak, Katarzyna Aleksenko, Svetlana S. Nosenko, Sergey V. Karandashev, Vasily K. Jarosz, Maciej Timerbaev, Andrei R. Anal Bioanal Chem Research Paper 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. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1007/s00216-017-0186-0) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2017-01-23 2017 /pmc/articles/PMC5352744/ /pubmed/28116493 http://dx.doi.org/10.1007/s00216-017-0186-0 Text en © The Author(s) 2017 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. |
spellingShingle | Research Paper Foteeva, Lidia S. Matczuk, Magdalena Pawlak, Katarzyna Aleksenko, Svetlana S. Nosenko, Sergey V. Karandashev, Vasily K. Jarosz, Maciej Timerbaev, Andrei R. Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions |
title | Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions |
title_full | Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions |
title_fullStr | Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions |
title_full_unstemmed | Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions |
title_short | Combination of ICP-MS, capillary electrophoresis, and their hyphenation for probing Ru(III) metallodrug–DNA interactions |
title_sort | combination of icp-ms, capillary electrophoresis, and their hyphenation for probing ru(iii) metallodrug–dna interactions |
topic | Research Paper |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5352744/ https://www.ncbi.nlm.nih.gov/pubmed/28116493 http://dx.doi.org/10.1007/s00216-017-0186-0 |
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