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This concise, self-contained and cohesive book focuses on commonly used and recently developed methods for designing and analyzing high-throughput screening (HTS) experiments from a statistically sound basis. Combining ideas from biology, computing and statistics, the author explains experimental designs and analytic methods that are amenable to rigorous analysis and interpretation of RNAi HTS experiments. The opening chapters are carefully presented to be accessible both to biologists with training only in basic statistics and to computational scientists and statisticians with basic biological knowledge. Biologists will see how new experiment designs and rudimentary data-handling strategies for RNAi HTS experiments can improve their results, whereas analysts will learn how to apply recently developed statistical methods to interpret HTS experiments.
Featuring new screening technologies as well as many well established methods, this book offers comprehensive treatment of the activities directly related to High Throughput Screening (HTS), such as compound library management, data handling, and robotics.
In High Throughput Screening, leading scientists and researchers expert in molecular discovery explain the diverse technologies and key techniques used in HTS and demonstrate how they can be applied generically. Writing to create precisely the introductory guidebook they wish had been available when they started in HTS, these expert seasoned authors illuminate the HTS process with richly detailed tutorials on the biological techniques involved, the management of compound libraries, and the automation and engineering approaches needed. Extensive discussions provide readers with all those key elements of pharmacology, molecular biology, enzymology, and biochemistry that will ensure the identification of suitable targets and screens, and detail the technology necessary to mine millions of data points for meaningful knowledge.
"High Throughput Screening (HTS) is one of several hit identification approaches that are part of a developing and evolving toolbox for the discovery of pharmaceutical start points. HTS remains one of the most successful approaches, and therefore an important foundation of drug discovery. In High Throughput Screening: Methods, Techniques and Applications, leading industrial and academic experts in screening and drug discovery explain key technologies and methods while demonstrating how they can be applied to successful hit identification. Describing both traditional and emerging methods in detail, this book provides an overview of these methods to the reader that will serve both those new to the field and expert scientists alike. High Throughput Screening: Methods, Techniques and Applications provides readers with an outline of key elements in the areas of assay development, detailed descriptions of a range of both biochemical and cell-based screening methodologies and strategies, as well as highlighting important steps in data analysis. By describing the basic principles of methods commonly used in HTS, High Throughput Screening: Methods, Techniques and Applications provides an illuminating introduction to HTS, capturing established good practice within the field, thereby imparting both the industrial and academic researcher with the knowledge required to work effectively in both today's and the hit identification laboratories of the future"--
As the use of high-throughput screening expands and creates more interest in the academic community, the need for detailed reference materials becomes ever more pressing. Cell-Based Assays for High-Throughput Screening: Methods and Protocols aims to fill an important part of this need by providing an easily accessible reference volume for cell-based phenotypic screening. Leading researchers in the field contribute state-of-the-art methods with actionable protocols covering four major areas of study: model biological systems, screening modalities and assay systems, detection technologies, and approaches to data analysis. Written in the highly successful Methods in Molecular BiologyTM series format, each chapter includes a brief introduction to the subject, lists of necessary materials and reagents, step-by-step laboratory protocols, and a Notes section detailing tips on troubleshooting and avoiding known pitfalls. Cutting-edge and easy-to-use, Cell-Based Assays for High-Throughput Screening: Methods and Protocols presents an overview of relevant approaches, enabling the direct application of existing methods to new discoveries while also inspiring researchers to approach their screening projects in a conceptually modular fashion, enhancing the power to discover through new combinations of existing approaches.
In the next couple of years the human genome will be fully sequenced. This will provide us with the sequence and overall function of all human genes as well as the complete genome for many micro-organisms. Subsequently it is hoped, by means of powerful bioinformatic tools, to determine the gene variants that contribute to various multifactorial diseases and genes that exist in certain infectious agents but not humans. As a consequence, this will allow us to define the most appropriate levels for drug intervention. It can be expected that the number of potential drug targets will increase, possibly by a factor of 10 or more. Nevertheless, sequencing the human genome or, for that matter, the genome of other species will only be the starting point for the understanding of their biological function. Structural genomics is a likely follow-up, combined with new techniques to validate the therapeutic relevance of such newly discovered targets. Accordingly, it can be expected that in the near future we will witness a substantial increase in novel putative targets for drugs. To address these new targets effectively, we require new approaches and innovative tools. At present, two alternative, yet complementary, techniques are employed: experimental high-throughput screening (HTS) of large compound libraries, increasingly provided by combinatorial chemistry, and computational methods for virtual screening and de novo design. As kind of status report on the maturity of virtual screening as a technique in drug design, the first workshop on new approaches in drug design and discovery was held in March 1999, at Schloß Rauischholzhausen, near Marburg in Germany. More than 80 scientists gathered and discussed their experience with the different techniques. The speakers were invited to summarize their contributions together with their impressions on the present applicability of their approach. Several of the speakers followed this request which is summarized in this publication.
Backed by leading authorities, this is a professional guide to successful compound screening in pharmaceutical research and chemical biology, including the chemoinformatic tools needed for correct data evaluation. Chapter authors from leading pharmaceutical companies as well as from Harvard University discuss such factors as chemical genetics, binding, cell-based and biochemical assays, the efficient use of compound libraries and data mining using cell-based assay results. For both academics and professionals in the pharma and biotech industries working on small molecule screening.
Directed evolution comprises two distinct steps that are typically applied in an iterative fashion: (1) generating molecular diversity and (2) finding among the ensemble of mutant sequences those proteins that perform the desired fu- tion according to the specified criteria. In many ways, the second step is the most challenging. No matter how cleverly designed or diverse the starting library, without an effective screening strategy the ability to isolate useful clones is severely diminished. The best screens are (1) high throughput, to increase the likelihood that useful clones will be found; (2) sufficiently sen- tive (i. e. , good signal to noise) to allow the isolation of lower activity clones early in evolution; (3) sufficiently reproducible to allow one to find small improvements; (4) robust, which means that the signal afforded by active clones is not dependent on difficult-to-control environmental variables; and, most importantly, (5) sensitive to the desired function. Regarding this last point, almost anyone who has attempted a directed evolution experiment has learned firsthand the truth of the dictum “you get what you screen for. ” The protocols in Directed Enzyme Evolution describe a series of detailed p- cedures of proven utility for directed evolution purposes. The volume begins with several selection strategies for enzyme evolution and continues with assay methods that can be used to screen enzyme libraries. Genetic selections offer the advantage that functional proteins can be isolated from very large libraries s- ply by growing a population of cells under selective conditions.
With the most comprehensive and up-to-date overview of structure-based drug discovery covering both experimental and computational approaches, Structural Biology in Drug Discovery: Methods, Techniques, and Practices describes principles, methods, applications, and emerging paradigms of structural biology as a tool for more efficient drug development. Coverage includes successful examples, academic and industry insights, novel concepts, and advances in a rapidly evolving field. The combined chapters, by authors writing from the frontlines of structural biology and drug discovery, give readers a valuable reference and resource that: Presents the benefits, limitations, and potentiality of major techniques in the field such as X-ray crystallography, NMR, neutron crystallography, cryo-EM, mass spectrometry and other biophysical techniques, and computational structural biology Includes detailed chapters on druggability, allostery, complementary use of thermodynamic and kinetic information, and powerful approaches such as structural chemogenomics and fragment-based drug design Emphasizes the need for the in-depth biophysical characterization of protein targets as well as of therapeutic proteins, and for a thorough quality assessment of experimental structures Illustrates advances in the field of established therapeutic targets like kinases, serine proteinases, GPCRs, and epigenetic proteins, and of more challenging ones like protein-protein interactions and intrinsically disordered proteins