Determination of Binding Constant of DNA-binding Drug to Target DNA by Surface Plasmon Resonance Biosensor Technology

Abstract

The experimental determination of the binding constant of a drug for its target molecule is of considerable importance. It is a basic experimental parameter in a variety of studies, such as the prediction of drug efficiency, or in the pharmacokinetic drug interaction. DNA-binding drugs have been reported to be able to interfere in a sequence dependent manner with biological functions such as topoisomerase activity, restriction of enzyme cleavage of DNA, protein-DNA interactions and the activity of transcription factors, leading to alteration of gene expression. This effect could have important practical application in the experimental therapy of human pathologies, including neoplastic diseases and viral, or microbial infections. The assessment of the biological activity of DNA-binding drugs by polymerase chain reaction, footprinting, gel retardation and in vitro transcription studies was recently reported. However, most of these techniques are steady-state methodologies and therefore are not suitable for an easy determination of the binding activity of DNA-binding drugs to target DNA and the stability of drugs-DNA complexes. Direct real-time observation and measurement of the interaction between DNA-binding drug and target DNA sequence is a subject of interest for drug discovery and development. The recent development of biosensors, based on surface plasmon resonance (SPR) technology, enables monitoring of a variety of biospecific interactions of DNA-binding drugs with target DNA elements in real-time. The present review is designed to indicate the theoretical background of SPR-based biosensor technology as well as to present the great variety of measurements and modes of interaction kinetics that can be performed with these techniques. In addition, some of the most recent studies in determining the binding constant and stoichiometry of DNA-binding drugs to target DNA with SPR technology are reviewed and the available theoretical aspects necessary for the comprehension of the experiments are provided.

Survey of the year 2005 commercial optical biosensor literature

“…O'Kennedy et al [12] described the technology's use for detecting microorganisms in water and food; Aguilar and Small [1] summarized the biosensor's recent contribution to understanding b-amyloid interactions and fibril formation related to Alzheimer's disease; Huber [8] outlined a biosensor-based strategy to streamline lead optimization in drug discovery; Swanson [17] demonstrated how the biosensor can be used in the development of therapeutic antibodies; and Choulier et al [5] and Lin et al [9] highlighted the biosensor's impact in characterizing viral proteins and small molecule/oligonucleotide interactions, respectively.…”

19F NMR applications in chemical biology

“…DNA has long been a target for the development of drug molecules [76] and consequently considerable effort has been directed towards developing analytical techniques to investigate the binding of small molecules to specific DNA targets. At present molecules that can stabilise DNA G-quadruplexes (quartet of four guanines) and thus act inhibitors of the telomerase enzyme are attracting considerable attention due to their potential anti-cancer properties [77].…”

Development of Surface Plasmon Resonance (SPR) Based Immuno-Sensing System for Detection of Fungal Teliospores of Karnal Bunt (Tilletia indica), a Quarantined Disease of Wheat

“…SPR techniques involve the excitation of surface plasmons using a light source at planar surfaces. These techniques have been used extensively to characterize binding reactions without labeling requirements [13]. This equipment is based on the Kretschmann configuration, which is also the most commonly-used configuration in SPR.…”