Monitoring, and estimation of kinetic data, by piezoelectric impedance analysis, of the binding of the anticancer drug mitoxantrone to surface-immobilized DNA

Abstract

A new method of monitoring drug binding to DNA, in real-time, by means of the piezoelectric quartz-crystal-impedance (PQCI) technique, is proposed. The method was used to monitor the binding of an anticancer drug, mitoxantrone (MX), to double-stranded calf-thymus DNA covalently immobilized on the thioglycollic acid-modified gold electrode surface of a quartz crystal. Optimum experimental conditions for the immobilization were established. The DNA-anchored piezoelectric sensor was in contact with MX solution. The time courses of the resonant frequency and the equivalent circuit characteristics of the sensor were also obtained during the study of DNA-drug binding. On the basis of the analysis of the multidimensional information provided by the PQCI technique the observed frequency decrease was mainly ascribed to the increase in the mass of the sensor surface resulting from the binding. The kinetics of the binding process were studied quantitatively by monitoring the frequency change with time and a piezoelectric response model for the binding was derived theoretically. The experimental data were fitted to the model and the binding and dissociation rate constants and the binding equilibrium constant were estimated to be 66.0+/-0.1 mol(-1) L s(-1), 1.4+/-0.1x10(-4) s(-1), and 4.71+/-0.07x10(5) mol(-1) L, respectively, at 37 degrees C.

A survey of the 2001 to 2005 quartz crystal microbalance biosensor literature: applications of acoustic physics to the analysis of biomolecular interactions

“…Piezoelectric nucleic acid sensors have been usefully applied to the investigation of various cancer drugs including anti-tumour antibiotics such as mitoxantrone (Mao et al, 2002), mitomycin and bleomycin (Tian et al, 2004), nogalamycin (Pope et al, 2001) and bleomycin in comparison to other analytes expected to bind to DNA (such as cisplatin and two intercalators) (Fucassi et al, 2001). Both Pope et al (2001) and Yau et al (2002) compared groove binders, namely Berenil and Hoechst 3342, respectively, to other cancer drugs using the QCM technique.…”

Biosensors and nanobiosensors for therapeutic drug and response monitoring

“…Surface plasmon resonance (SPR) sensing, for example, is currently the most common technique for monitoring small molecule interactions with receptors, 30 and other biosensing techniques are also suited for this purpose. Hence, SPR along with electrochemical sensors, [31][32][33] fibre-optic sensing 34 and piezoelectric impedance analysis 35 were identified early on as potential platforms for therapeutic drug monitoring. Already, SPR sensing, 36,37 LSPR sensing, 36,38 fluorescence based sensing, 11,39,40 quartz crystal microbalance (QCM), 41,42 paper-spray mass spectrometry [43][44][45] and Raman spectroscopy [46][47][48][49] have been applied to therapeutic drug monitoring in the literature.…”

“…It is nonetheless possible to detect small molecules with SPR 30 and this has led to the widespread implementation of SPR sensing in pharmaceutical laboratories for ligand screening assays. 28,67 More precisely, to circumvent the issue of lower sensitivity, indirect techniques such as competition assays 36 or secondary detection with antibodies or functionalized nanoparticles 35 can be utilized to increase the response in SPR, and many (nano)biosensors for therapeutic drug monitoring to date have been based on competition assays.…”

Estimation of Kinetics Parameters for the Adsorption of Human Serum Albumin onto Hydroxyapatite-modified Silver Electrodes by Piezoelectric Quartz Crystal Impedance Analysis

“…The quality of the fitting can be evaluated by the relative sum of the residual square, qr, which was defined in our previous report. 28 From the above adsorption model, the experimental data were fitted to Eq. (12).…”