Utilizing inherent fluorescence of therapeutics to analyze real-time uptake and multi-parametric effector kinetics

Autoren

Benjamin Wiench
Tolga Eichhorn
Bernhard Korn
Malte Paulsen
Thomas Efferth

DOI

10.1016/j.ymeth.2012.01.007

ISSN

1046-2023

Ausgabe der Veröffentlichung

3

Zeitschrift

Methods

Sprache

en

Paginierung

376 - 382

Datum der Veröffentlichung

2012

Status

Published

Herausgeber

Elsevier BV

Herausgeber URL

http://dx.doi.org/10.1016/j.ymeth.2012.01.007

Datum der Datenerfassung

2023

Titel

Utilizing inherent fluorescence of therapeutics to analyze real-time uptake and multi-parametric effector kinetics

Ausgabe der Zeitschrift

57

Data source: Crossref

Abstract

The precise detection of pharmaceutical drug uptake and knowledge of a drug's efficacy at the single-cell level is crucial for understanding a compound's performance. Many pharmaceutical drugs, like the model substances Doxorubicin, Mitoxantrone or Irinotecan, have a distinctive natural fluorescence that can be readily exploited for research purposes. Utilizing this respective natural fluorescence, we propose a method analyzing simultaneously in real-time the efficiency, effects and the associated kinetics of compound-uptake and efflux in mammalian cells by flow cytometry. We show that real-time flow cytometric quantification of compound-uptake is reliably measured and that analyzing their respective uptake kinetic provides additional valuable information which can be used for improving drug dosage and delivery. Exploiting the native fluorescence of natural compounds is clearly advantageous compared to the usage of non-related fluorescent uptake-reporter substances, possibly yielding in unphysiological data. Flow cytometric analysis allows live-dye based multi-parametric high-throughput screening of pharmaceutical compound activity, improving cytotoxicity testing by combining several assays into a single, high resolution readout. This approach can be a useful tool identifying potential inhibitors for multiple drug resistance (MDR), representing a major challenge to the targeted treatment of various diseases.

Addresses

Institute of Pharmacy and Biochemistry, Department of Pharmaceutical Biology, Johannes Gutenberg University, Staudinger Weg 5, 55128 Mainz, Germany.

Autoren

Benjamin Wiench
Tolga Eichhorn
Bernhard Korn
Malte Paulsen
Thomas Efferth
Thomas Efferth

DOI

10.1016/j.ymeth.2012.01.007

eISSN

1095-9130

Externe Identifier

PubMed Identifier: 22326879

Open access

false

ISSN

1046-2023

Ausgabe der Veröffentlichung

3

Zeitschrift

Methods (San Diego, Calif.)

Schlüsselwörter

Cell Line, Tumor
Humans
Reactive Oxygen Species
Mitoxantrone
Camptothecin
Fluoresceins
Doxorubicin
Antineoplastic Agents
Flow Cytometry
Drug Resistance, Multiple
Cell Survival
Biological Transport
Drug Resistance, Neoplasm
Kinetics
Fluorescence
Single-Cell Analysis
Irinotecan

Sprache

eng

Medium

Print-Electronic

Online publication date

2012

Paginierung

376 - 382

Datum der Veröffentlichung

2012

Status

Published

Datum der Datenerfassung

2012

Titel

Utilizing inherent fluorescence of therapeutics to analyze real-time uptake and multi-parametric effector kinetics.

Sub types

Research Support, Non-U.S. Gov't
Journal Article

Ausgabe der Zeitschrift

57

Data source: Europe PubMed Central

Abstract

The precise detection of pharmaceutical drug uptake and knowledge of a drug's efficacy at the single-cell level is crucial for understanding a compound's performance. Many pharmaceutical drugs, like the model substances Doxorubicin, Mitoxantrone or Irinotecan, have a distinctive natural fluorescence that can be readily exploited for research purposes. Utilizing this respective natural fluorescence, we propose a method analyzing simultaneously in real-time the efficiency, effects and the associated kinetics of compound-uptake and efflux in mammalian cells by flow cytometry. We show that real-time flow cytometric quantification of compound-uptake is reliably measured and that analyzing their respective uptake kinetic provides additional valuable information which can be used for improving drug dosage and delivery. Exploiting the native fluorescence of natural compounds is clearly advantageous compared to the usage of non-related fluorescent uptake-reporter substances, possibly yielding in unphysiological data. Flow cytometric analysis allows live-dye based multi-parametric high-throughput screening of pharmaceutical compound activity, improving cytotoxicity testing by combining several assays into a single, high resolution readout. This approach can be a useful tool identifying potential inhibitors for multiple drug resistance (MDR), representing a major challenge to the targeted treatment of various diseases.

Date of acceptance

2012

Autoren

Benjamin Wiench
Tolga Eichhorn
Bernhard Korn
Malte Paulsen
Thomas Efferth

Autoren-URL

https://www.ncbi.nlm.nih.gov/pubmed/22326879

DOI

10.1016/j.ymeth.2012.01.007

eISSN

1095-9130

Ausgabe der Veröffentlichung

3

Zeitschrift

Methods

Schlüsselwörter

Antineoplastic Agents
Biological Transport
Camptothecin
Cell Line, Tumor
Cell Survival
Doxorubicin
Drug Resistance, Multiple
Drug Resistance, Neoplasm
Flow Cytometry
Fluoresceins
Fluorescence
Humans
Irinotecan
Kinetics
Mitoxantrone
Reactive Oxygen Species
Single-Cell Analysis

Sprache

eng

Country

United States

Paginierung

376 - 382

PII

S1046-2023(12)00023-0

Datum der Veröffentlichung

2012

Status

Published

Datum, an dem der Datensatz öffentlich gemacht wurde

2013

Titel

Utilizing inherent fluorescence of therapeutics to analyze real-time uptake and multi-parametric effector kinetics.

Sub types

Journal Article
Research Support, Non-U.S. Gov't

Ausgabe der Zeitschrift

57

Data source: PubMed

Utilizing inherent fluorescence of therapeutics to analyze real-time uptake and multi-parametric effector kinetics

Tools