Direct optical detection in fragment-based screening

Label-free biosensors based on direct optical detection principles are widely used in many different fields of research. Currently the higher level of automation and the increasing throughput of this technology are stimulating the interest of pharmaceutical companies. The information gained with label-free biosensors can be extremely valuable during the drug design process, particularly in combination with complementary techniques, including NMR, mass spectrometry and X-ray crystallography. In this article we focus on the advantages of direct optical biosensors especially in the field of fragment-based drug design, which is a widely used and extremely promising concept. Furthermore, we present optical biosensors as versatile tools for fragment-based screening and the future drug design process.     

基于纳米粒子的光学细胞传感器在癌细胞研究中的应用

作为一种极为灵敏、快速的新型生物检测技术,光学细胞传感器在生物医学领域的研究应用备受关注,成为当今生物分析化学领域的研究前沿和热点.它是以细胞作为传感元件来研究信号识别、传导和指示的过程,在毒性物质、病原体等外界条件作用下,研究细胞中活性分子及其生理条件的变化,通过光学信号的变化定量分析细胞膜表面分子、胞内酶分子及微环...     

Chip-based high-throughput screening of herbal medicines

At present, high-throughput screening (HTS) programs in drug discovery rely mainly on compound libraries from combinational chemistry. Similarly, natural flora has been used as a prominent origin for new and potent herbal drugs. Herbal medicines have been used worldwide for thousands of years to cure many diseases. As such, herbal secondary metabolites show a remarkable structural diversity that supplements chemically synthesized compound analogs in drug discovery screening. Unfortunately, there is often a considerable deterioration in the quality of herbal drugs in such screening programs as there are time-consuming manual processes involved in the isolation of active ingredients from the highly complex mixtures of herbal plant products. The quality and quantity of herbal samples are critical for the success of HTS programs. In the recent past, there have been substantial improvements in HTS due to the miniaturization and integration of microchip (e.g., Herbochip(?), DNA chip, protein chip, cell chip, etc.)-based technologies so as to design herbal drugs that compete with synthetic drug analogs. Here we will review various technologies used for HTS of herbal medicines. Finally, we will summarize our efforts to develop a novel chip-based HTS assay to explore the antioxidant and radioprotective properties of herbal plants.     

Integration of virtual and high throughput screening in lead discovery settings

In the last decade mass screening strategies became the main source of leads in drug discovery settings. Although high throughput (HTS) and virtual screening (VS) realize the same concept the different nature of these lead discovery strategies (experimental vs theoretical) results that they are typically applied separately. The majority of drug leads are still identified by hit-to-lead optimization of screening hits. Structural information on the target as well as on bound ligands, however, make structure-based and ligand-based virtual screening available for the identification of alternative chemical starting points. Although, the two techniques have rarely been used together on the same target, here we review the existing prominent studies on their true integration. Various approaches have been shown to apply the combination of HTS and VS and to better use them in lead generation. Although several attempts on their integration have only been considered at a conceptual level, there are numerous applications underlining its relevance that early-stage pharmaceutical drug research could benefit from a combined approach.     

Biosensors in Drug Discovery and Drug Analysis

Abstract

Biosensors are, by definition, sensing devices comprising a biological component (enzyme, antibody, animal or plant cell, oligonucleotide, lipid, microorganisms, etc.) intimately connected to a physical transducer (electrode, optical fiber, vibrating quartz, etc.). This dual configuration permits a quantitative study of the interaction between a drug compound and an immobilized biocomponent. Ideally, biosensors should be readily implemented and allow for low reagent and energy consumption. Enzyme‐based biosensors can be applied in the pharmaceutical industry for monitoring chemical parameters in the production process (in bioreactors). Affinity biosensors are suitable for high‐throughput screening of bioprocess‐produced antibodies and for candidate drug screening. They are suitable for selective and sensitive immunoassays in clinical laboratories and for decentralized detection of drug residues. Enzyme‐based biosensors may be used in hospitals for bedside drug testing, emergency control, in patient treatment control (anticancer therapy) etc. Current research efforts are focused on proteins, tissues, or living cells immobilized in microfabricated configurations for high‐throughput drug screening and discovery. Such devices can comprise several different microelectronic sensors and biosensors sensitive, for example, to pH, temperature, impedance, dissolved oxygen, etc. for a multiparametric monitoring. Of equal new interest are the oligonucleotide‐immobilized biosensors for interactions studies between a surface linked DNA and the target drug or for hybridisation studies. This short review summarizes several recent trends dedicated to the development and application of biosensors in the pharmaceutical arena.     

Natural and artificial ion channels for biosensing platforms

The single-molecule selectivity and specificity of the binding process together with the expected intrinsic gain factor obtained when utilizing flow through a channel have attracted the attention of analytical chemists for two decades. Sensitive and selective ion channel biosensors for high-throughput screening are having an increasing impact on modern medical care, drug screening, environmental monitoring, food safety, and biowarefare control. Even virus antigens can be detected by ion channel biosensors. The study of ion channels and other transmembrane proteins is expected to lead to the development of new medications and therapies for a wide range of illnesses. From the first attempts to use membrane proteins as the receptive part of a sensor, ion channels have been engineered as chemical sensors. Several other types of peptidic or nonpeptidic channels have been investigated. Various gating mechanisms have been implemented in their pores. Three technical problems had to be solved to achieve practical biosensors based on ion channels: the fabrication of stable lipid bilayer membranes, the incorporation of a receptor into such a structure, and the marriage of the modified membrane to a transducer. The current status of these three areas of research, together with typical applications of ion-channel biosensors, are discussed in this review.     

层层自组装聚电解质膜光纤传感器的制备和应用

在过去几十年中,光纤的应用已经渗透到多个学科领域。光纤的抗电磁干扰、可远程监控、多重监测、体积小及质量轻等特点,使其在传感器研究领域备受关注。聚电解质层层自组装膜构建的光纤传感器自2000年诞生以来,已快速发展成为传感器领域新的研究热点。该类光纤传感器在微量物质的监测方面具有广泛的应用前景。本文从光纤和光纤传感器优点出发,总结了基于层层自组装多层膜的光纤传感器种类、性能、检测原理以及相应的光纤结构和自组装材料;进而结合作者已做的相关工作,论述了在光纤基底上的聚电解质层层自组装及基于自组装膜的光纤传感器的测试;重点综述了近十年层层自组装膜的光纤pH传感器、湿度传感器、气体传感器、生物传感器及其他类型的光纤传感器的制备与应用,并展望了今后聚电解质层层自组装多层膜光纤传感器的发展。     

Conjugated Polyelectrolytes as New Platforms for Drug Screening

In recent years, conjugated polyelectrolytes (CPEs) have attracted increasing attention for their applications in highly sensitive biosensors by taking advantage of their unique optical amplification properties. In comparison to previous applications tailored for highly sensitive biomacromolecule detection, this Focus Review highlights recent research efforts in the development of water‐soluble CPEs as a new class of optical platforms for the screening of potential drugs. Three types of biological targets for the search of small‐molecule active drugs are described: nucleic acids, enzymes, and RNA–protein complexes. Future research directions for drug screening based on CPEs are also presented.     

Recent advances in nucleic acid amplification-based optical biosensors for disease diagnosisEI北大核心CSCD

Optical biosensors have been widely used in the detection of biomarkers due to their advantages of simple operationquick responsehigh sensitivity and visualization. When constructing optical biosensors nucleic acid amplification technology can be used to improve the analytical performance of optical biosensor which can further realize the highly sensitive detection of biomarkers and provide more accurate information for disease diagnosis. In this reviewrecent advances in nucleic acid amplification-based optical biosensors for disease diagnosis were reviewed the possible problems may exist in practical applications and future development trends were proposed. © 2022, Youke Publishing Co.,Ltd. All rights reserved.     

K-Screen: a free application for high throughput screening data analysis, visualization, and laboratory information management

High throughput screening (HTS) has emerged as an important technique for allowing researchers to rapidly profile very large numbers of chemicals against drug targets. As recent and future advances make HTS cheaper to perform on even larger scales, the amount of data that has to be processed, analyzed, and searched will only grow larger in size and harder for researchers to manually sift through. It is therefore an unavoidable requirement that institutions utilizing HTS technology will need to begin looking for effective solutions in the maturing area of laboratory information management systems like many other types of labs have already done. K-Screen is one such solution. Our initial goal with K-Screen was to have an integrated application environment that supported data analysis, management, and presentation so we could efficiently perform client requested screens and searches as well as generate detailed reports on the results of those. Previously, we had attempted but failed to locate an existing software suite that sufficiently addressed all our requirements. K-Screen is a web accessible application that offers the ability to host a large chemical structure library, process and store single-dose (primary) and dose response (secondary) screening data, perform searches based on screening results, plate coordinates, and structure, substructure and structure similarity. It uses heat maps and histograms to visualize screen or plate level statistics. Interfaces to external searches against PubChem and ZINC databases are also provided. We feel that these features make K-Screen a practical and effective alternative to other commercial or academic HTS LIMS systems.     

High throughput screening in agrochemical research

The demand for new herbicides, insecticides and fungicides led to a steady increase in the number of compounds being tested to find novel market products. To keep pace with the rising workload, high throughput screening (HTS) technologies have been introduced. In agrochemical research miniaturised in vivo tests on whole real target organisms are now possible and are an integral part of the screening cascade. A complementary target based in vitro HTS has also been established in agrochemical research. Target based HTS allows a directed approach towards untouched market shares by novel modes of action. Selection of the best suited targets is the most crucial issue in this approach. Genomic methods thereby deliver many essential genes as candidate targets. Consideration of further criteria such as druggability notably narrows down the number of promising targets. Though target to hit to lead progression still is as in pharmaceutical research a complex and therefore risky process, the implementation of novel bioscience technologies has entailed the transition to an integrated innovative agrochemical research perspective.     

Modern drug discovery technologies: opportunities and challenges in lead discovery

The identification of promising hits and the generation of high quality leads are crucial steps in the early stages of drug discovery projects. The definition and assessment of both chemical and biological space have revitalized the screening process model and emphasized the importance of exploring the intrinsic complementary nature of classical and modern methods in drug research. In this context, the widespread use of combinatorial chemistry and sophisticated screening methods for the discovery of lead compounds has created a large demand for small organic molecules that act on specific drug targets. Modern drug discovery involves the employment of a wide variety of technologies and expertise in multidisciplinary research teams. The synergistic effects between experimental and computational approaches on the selection and optimization of bioactive compounds emphasize the importance of the integration of advanced technologies in drug discovery programs. These technologies (VS, HTS, SBDD, LBDD, QSAR, and so on) are complementary in the sense that they have mutual goals, thereby the combination of both empirical and in silico efforts is feasible at many different levels of lead optimization and new chemical entity (NCE) discovery. This paper provides a brief perspective on the evolution and use of key drug design technologies, highlighting opportunities and challenges.     

Fluorescence polarization immunoassays and related methods for simple, high-throughput screening of small molecules

Fluorescence polarization immunoassay (FPIA) is a homogeneous (without separation) competitive immunoassay method based on the increase in fluorescence polarization (FP) of fluorescent-labeled small antigens when bound by specific antibody. The minimum detectable quantity of FPIAs with fluorescein label (about 0.1 ng analyte) is comparable with chromatography and ELISA methods, although this may be limited by sample matrix interference. Because of its simplicity and speed, FPIA is readily automated and therefore suitable for high-throughput screening (HTS) in a variety of application areas. Systems that involve binding of ligands to receptor proteins are also susceptible to analysis by analogous FP methods employing fluorescent-labeled ligand and HTS applications have been developed, notably for use in candidate drug screening.     

Parallel and multiplexed bead-based assays and encoding strategies

Advances in high throughput screening (HTS), together with the rapid progress in combinatorial chemistry, genomic and proteomic sciences have dramatically stimulated the development of a variety tools to enable the drug discovery process to become more efficient. Major future challenges in HTS include obtaining high density and good quality data based on assays that are rapid, reliable, inexpensive, sensitive, simple and miniaturised. This paper reviews the development and role of bead-based assays for HTS including DNA and single nucleotide polymorphism (SNP) assays, particularly from a multiplex perspective and evaluating the recent advances in bead-based arrays. The encoding strategies that are commonly used in bead-based assays are highlighted, while the importance of magnetic beads in genomic and proteomic purifications is discussed. In conclusion, bead-based assays offer a powerful promising approach for many aspects of drug discovery.     

Optical biosensors based on Prussian Blue films

Optical biosensing schemes based on enzymatically modified inorganic/organic transparent films predominately composed of Prussian Blue are demonstrated. The composite film, which is non-electrochemically deposited on a non-conducting support. is used as an optical transducer for flow-through biosensors based on hydrolases and oxidases. Urease and glucose oxidase are utilized as model enzymes. Action of the urea biosensor is based on optical pH sensitivity of Prussian Blue indicator. The glucose biosensor is acting as first-generation optical biosensor based on in situ generated Prussian White transducer for hydrogen peroxide. These simple, single-pass transmission optical biosensors exhibit sensitivity in the millimolar range of concentration. The biosensors are very stable owing to presence of a poly(pyrrolylbenzoic acid) network in the composite material. This organic polymer plays a dual role as a binding agent for inorganic material and as a functionalized support for strong covalent immobilization of enzyme molecules.     

High throughput screening informatics

High throughput screening (HTS), an industrial effort to leverage developments in the areas of modern robotics, data analysis and control software, liquid handling devices, and sensitive detectors, has played a pivotal role in the drug discovery process, allowing researchers to efficiently screen millions of compounds to identify tractable small molecule modulators of a given biological process or disease state and advance them into high quality leads. As HTS throughput has significantly increased the volume, complexity, and information content of datasets, lead discovery research demands a clear corporate strategy for scientific computing and subsequent establishment of robust enterprise-wide (usually global) informatics platforms, which enable complicated HTS work flows, facilitate HTS data mining, and drive effective decision-making. The purpose of this review is, from the data analysis and handling perspective, to examine key elements in HTS operations and some essential data-related activities supporting or interfacing the screening process, and outline properties that various enabling software should have. Additionally, some general advice for corporate managers with system procurement responsibilities is offered.     

High throughput screening for neurodegeneration and complex disease phenotypes

High throughput screening (HTS) for complex diseases is challenging. This stems from the fact that complex phenotypes are difficult to adapt to rapid, high throughput assays. We describe the recent development of high throughput and high-content screens (HCS) for neurodegenerative diseases, with a focus on inherited neurodegenerative disorders, such as Huntington's disease. We describe, among others, HTS assays based on protein aggregation, neuronal death, caspase activation and mutant protein clearance. Furthermore, we describe high-content screens that are being used to prioritize hits identified in such HTS assays. These assays and screening approaches should accelerate drug discovery for neurodegenerative disorders and guide the development of screening approaches for other complex disease phenotypes.     

Whole cell strategies based on lux genes for high throughput applications toward new antimicrobials

The discovery/development of novel drug candidates has witnessed dramatic changes over the last two decades. Old methods to identify lead compounds are not suitable to screen wide libraries generated by combinatorial chemistry techniques. High throughput screening (HTS) has become irreplaceable and hundreds of different approaches have been described. Assays based on purified components are flanked by whole cell-based assays, in which reporter genes are used to monitor, directly or indirectly, the influence of a chemical over the metabolism of living cells. The most convenient and widely used reporters for real-time measurements are luciferases, light emitting enzymes from evolutionarily distant organisms. Autofluorescent proteins have been also extensively employed, but proved to be more suitable for end-point measurements, in situ applications - such as the localization of fusion proteins in specific subcellular compartments - or environmental studies on microbial populations. The trend toward miniaturization and the technical advances in detection and liquid handling systems will allow to reach an ultra high throughput screening (uHTS), with 100,000 of compounds routinely screened each day. Here we show how similar approaches may be applied also to the search for new and potent antimicrobial agents.     

高通量药物筛选现代检测技术研究进展

高通量药物筛选是发现创新药物的重要技术途径.高通量筛选结果必须通过适当的检测方法才能反映出来,检测技术是实现高通量药物筛选的基础.本文综述了近年来有关光学分析、色谱分析、热分析、电化学分析、质谱、核磁共振等现代检测技术在高通量药物筛选研究中的进展.     

High throughput screening technology and the small molecules modulating aging related signals

Aging and its related diseases are severe issues in modern society. Many efforts have been made to understand the mechanisms of aging and to find the ways to prevent age-related diseases. Identifying the compounds targeting aging-related signals is a challenging work because there are so many proteins and signals involved. Recently, alone with the progresses in high throughput screening (HTS) technology, increasing numbers of small molecules targeting aging-related pathologic processes have been identified. In this review, we introduce the basic workflow, classification and assay strategies of HTS technology, and sort out known small molecules identified via HTS technology by their roles in aging related diseases, such as neural degenerative diseases, diabetes and tumors. Given the fact that application of HTS on aging research is still at an early stage, we also summarize the cellular mechanisms about aging process, paralleled with the compounds which can modulate the functions of proteins important for aging signals. Finally, we briefly discuss some advanced HTS technologies for their potent applications on the discovery of anti-aging compounds. The main purpose of this review is to provide updated and useful information to those who are interested in pharmacology and HTS technology, but not familiar with aging biology, or vice versa.