Immunotoxin Methods and Protocols
Immunotoxin Methods and Protocols

Author: Walter A. Hall

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 305

ISBN-13: 1592591140

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Immunotoxins represent a new class of human therapeutics that have widespread applications and a potential that has not yet been fully recognized since they were first conceived of by Paul Ehrlich in 1906. The majority of advances in the development and implementation of immunotoxins has occurred over the last 20 years. The reasons for this use of immunotoxins in basic science and clinical research are the powerful concurrent advances in genetic engineering and receptor physiology. Recombinant technology has allowed investigators to produce sufficient quantities of a homogeneous c- pound that allows clinical trials to be performed. The identification of specific receptors on malignant cell types has enabled scientists to generate immunotoxins that have had positive results in clinical trials. As more cellular targets are identified in coming years, additional trials will be conducted in different disease states affecting still larger patient populations. Modulation of the immune system to decrease the humoral response to immunotoxins may improve their overall efficacy. As increasingly more effective compounds are generated, it will be necessary to decrease the local and systemic toxicity - sociated with these agents, and methods for doing so are presently being - veloped. The work presented in Immunotoxin Methods and Protocols focuses on three specific areas of immunotoxin investigation that are being conducted by experts throughout the world. The first section describes the construction and development of a variety of immunotoxins.

Nuclease Methods and Protocols
Nuclease Methods and Protocols

Author: Catherine H. Schein

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 521

ISBN-13: 1592592333

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Nucleases, enzymes that restructure or degrade nucleic acid polymers, are vital to the control of every area of metabolism. They range from “housekeeping” enzymes with broad substrate ranges to extremely specific tools (1). Many types of nucleases are used in lab protocols, and their commercial and clinical uses are expanding. The purpose of Nuclease Methods and Protocols is to introduce the reader to some we- characterized protein nucleases, and the methods used to determine their activity, structure, interaction with other molecules, and physiological role. Each chapter begins with a mini-review on a specific nuclease or a nuclease-related theme. Although many chapters cover several topics, they were arbitrarily divided into five parts: Part I, “Characterizing Nuclease Activity,” includes protocols and assays to determine general (processive, distributive) or specific mechanisms. Methods to assay nuclease products, identify cloned nucleases, and determine their physiological role are also included here. Part II, “Inhibitors and Activators of Nucleases,” summarizes assays for measuring the effects of other proteins and small molecules. Many of these inhibitors have clinical relevance. Part III, “Relating Nuclease Structure and Function,” provides an overview of methods to determine or model the 3-D structure of nucleases and their complexes with substrates and inhibitors. A 3-D structure can greatly aid the rational design of nucleases and inhibitors for specific purposes. Part IV, “Nucleases in the Clinic,” summarizes assays and protocols suitable for use with t- sues and for nuclease based therapeutics.

Connexin Methods and Protocols
Connexin Methods and Protocols

Author: Roberto Bruzzone

Publisher: Springer Science & Business Media

Published: 2008-02-05

Total Pages: 495

ISBN-13: 1592590438

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Direct cell–cell communication is a common property of multicellular organisms that is achieved through membrane channels which are organized in gap junctions. The protein subunits of these intercellular channels, the connexins, form a multigene family that has been investigated in great detail in recent years. It has now become clear that, in different tissues, connexins speak several languages that control specific cellular functions. This progress has been made possible by the availability of new molecular tools and the improvement of basic techniques for the study of membrane channels, as well as by the use of genetic approaches to study protein function in vivo. More important, connexins have gained visibility because mutations in some connexin genes have been found to be linked to human genetic disorders. Connexin Methods and Protocols presents in detail a collection of te- niques currently used to study the cellular and molecular biology of connexins and their physiological properties. The field of gap junctions and connexin research has always been characterized by a multidisciplinary approach c- bining morphology, biochemistry, biophysics, and cellular and molecular biology. This book provides a series of cutting-edge protocols and includes a large spectrum of practical methods that are available to investigate the fu- tion of connexin channels. Connexin Methods and Protocols is divided into three main parts.

Capillary Electrophoresis of Nucleic Acids
Capillary Electrophoresis of Nucleic Acids

Author: Keith R. Mitchelson

Publisher: Springer Science & Business Media

Published: 2008-02-05

Total Pages: 481

ISBN-13: 1592590551

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The development of PCR, which enables extremely small amounts of DNA to be amplified, led to the rapid development of a multiplicity of a- lytical procedures that permit use of this new resource for the analysis of genetic variation and for the detection of disease-causing mutations. The advent of capillary electrophoresis (CE), with its power to separate and a- lyze very small amounts of DNA, has also stimulated researchers to develop analytical procedures for the CE format. The advantages of CE in terms of speed and reproducibility of analyses are manifold. Furthermore, the high s- sitivity of detection, and the ability to increase sample throughput with par- lel analysis, has led to the creation of a full range of analysis of DNA molecules, from modified DNA adducts and single-strand oligonucleotides through PCR-amplified DNA fragments and whole chromosomes. Capillary Elect- phoresis of Nucleic Acids focuses on analytical protocols that can be used for detection and analysis of mutations and modification, from precise DNA loci through entire genomes of organisms. Important practical considerations for CE, such as the choice of separation media, electrophoresis conditions, and the influence of buffer additives and dyes on DNA mobility, are discussed in several key chapters and within particular applications.

The ELISA Guidebook
The ELISA Guidebook

Author: John R. Crowther

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 426

ISBN-13: 1592590497

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John R. Crowther provides today's premier practical guide to the understanding and application of ELISA. Updating and greatly expanding his widely appreciated earlier publication, ELISA Theory and Practice (1995), this important work introduces chapters on such major new topics as checkerboard titrations, quality control of testing, kit production and control, novel monoclonal antibodies, validation of assays, statistical requirements for data examination, and epidemiological considerations. With its numerous worked examples, detailed instructions, and extensive illustrations, The ELISA Guidebook offers a powerful synthesis of all the basic concepts and practical experimental details investigators need to understand, develop, and apply the new ELISA methodology successfully in day-to-day basic and clinical research.

Bioconjugation
Bioconjugation

Author: Sam Massa

Publisher: Humana

Published: 2019-07-23

Total Pages: 317

ISBN-13: 9781493996537

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This book explores well-established and emerging conjugation strategies that are relevant for proteins used in the field of precision medicine, focusing on techniques that are suitable for antibodies, antibody-fragments such as Fabs, scFvs, or nanobodies, scaffold proteins such as FN3 or DARPin, peptides, or model proteins. Although centered on the development of bioconjugates rather than their application, most protocols also show the conjugation of the targeting vehicle to a diagnostic or therapeutic entity, with the end-product most often being an antibody-drug conjugate, an optical probe, a nanomedicine, or a radiopharmaceutical. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Bioconjugation: Methods and Protocols is an ideal guide for researchers looking toward precision medicine in order to expand the vital field of drug discovery.

Biostatistical Methods
Biostatistical Methods

Author: Stephen W. Looney

Publisher: Springer Science & Business Media

Published: 2008-02-03

Total Pages: 221

ISBN-13: 1592592422

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Leading biostatisticians and biomedical researchers describe many of the key techniques used to solve commonly occurring data analytic problems in molecular biology, and demonstrate how these methods can be used in the development of new markers for exposure to a risk factor or for disease outcomes. Major areas of application include microarray analysis, proteomic studies, image quantitation, genetic susceptibility and association, evaluation of new biomarkers, and power analysis and sample size.

Protein Sequencing Protocols
Protein Sequencing Protocols

Author: Bryan John Smith

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 489

ISBN-13: 1592593429

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Determination of the protein sequence is as important today as it was a half century ago, even though the techniques and purposes have changed over time. Mass spectrometry has continued its recent rapid development to find notable application in the characterization of small amounts of protein, for example, in the field of proteomics. The “traditional” chemical N-terminal sequencing is still of great value in quality assurance of the increasing number of biopharmaceuticals that are to be found in the clinic, checking processing events of recombinant proteins, and so on. It is joined in the armory of me- ods of protein analysis by such techniques as C-terminal sequencing and amino acid analysis. These methods are continually developing. The first edition of Protein Sequencing Protocols was a “snapshot” of methods in use in protein biochemistry laboratories at the time, and this, the second edition, is likewise. Methods have evolved in the intervening period, and the content of this book has similarly changed, the content of some chapters having been superceded and replaced by other approaches. Thus, in this edition, there is inclusion of approaches to validation of methods for quality assurance work, reflecting the current importance of biopharmaceuticals, and also a guide to further analysis of protein sequence information, acknowledging the importance of bioinformatics.

E. coli Gene Expression Protocols
E. coli Gene Expression Protocols

Author: Peter E. Vaillancourt

Publisher: Springer Science & Business Media

Published: 2008-02-02

Total Pages: 347

ISBN-13: 1592593011

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Peter E. Vaillancourt presents a collection of popular and emerging methodologies that take advantage of E. coli's ability to quickly and inexpensively express recombinant proteins. The authors focus on two areas of interest: the use of E. coli vectors and strains for production of pure, functional protein, and the use of E. coli as host for the functional screening of large collections of proteins and peptides. Among the cutting-edge techniques demonstrated are those for rapid high-level expression and purification of soluble and functional recombinant protein and those essential to functional genomics, proteomics, and protein engineering.

RT-PCR Protocols
RT-PCR Protocols

Author: Nicola King

Publisher: Springer Science & Business Media

Published: 2008-02-04

Total Pages: 370

ISBN-13: 159259283X

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Until the mid 1980s, the detection and quantification of a specific mRNA was a difficult task, usually only undertaken by a skilled molecular biologist. With the advent of PCR, it became possible to amplify specific mRNA, after first converting the mRNA to cDNA via reverse transcriptase. The arrival of this technique—termed reverse transcription-PCR (RT-PCR)—meant that mRNA suddenly became amenable to rapid and sensitive analysis, without the need for advanced training in molecular biology. This new accessibility of mRNA, which has been facilitated by the rapid accumulation of sequence data for human mRNAs, means that every biomedical researcher can now include measurement of specific mRNA expression as a routine component of his/her research plans. In view of the ubiquity of the use of standard RT-PCR, the main objective of RT-PCR Protocols is essentially to provide novel, useful applications of RT-PCR. These include some useful adaptations and applications that could be relevant to the wider research community who are already familiar with the basic RT-PCR protocol. For example, a variety of different adaptations are described that have been employed to obtain quantitative data from RT-PCR. Quantitative RT-PCR provides the ability to accurately measure changes/imb- ances in specific mRNA expression between normal and diseased tissues.