Basic Protein And Peptide Protocols

Basic Protein and Peptide Protocols offers an excellent collection of reproducible, step-by-step laboratory methods covering three major areas: (1) the quantitation and characterization of proteins, (2) the electrophoretic and blotting procedures used in protein isolation and characterization, and (3) the analysis of protein and peptide structure. THOUSANDS of labs are already using Basic Protein and Peptide Protocols-you should be too!

-- 145 concise, step-by-step protocols for the analysis of proteins and peptides -- Provides more than three times as many protocols than the competitors' manuals -- in the incomparable Methods in Molecular Biology "TM" style -- at an affordable price -- Combound in a large 7 x l0 inch format for easy benchtop use -- An indispensable reference for industry-based researchers, professors, and their students

Hans Neurath has written that this is the second golden era of enzymology {Protein Science [1994], vol. 3, pp. 1734—1739); he could with justice have been more general and referred to the second golden age of protein chemistry. The last two decades have seen enormous advances in our understanding of the structures and functions of pro teins arising on the one hand from improvements and developments in analytical techniques {see the companion volume, Basic Protein and Peptide Protocols, in this series) and on the other hand from the tech nologies of molecular genetics. Far from turning the focus away from protein science, the ability to isolate, analyze, and express genes has increased interest in proteins as gene products. Hence, many laborato ries are now getting involved in protein isolation for the first time, either as an essential adjunct to their work in molecular genetics or because of a curiosity to know more about the products of the genes that they have been studying. Protein Purification Protocols is aimed mainly at these newcom ers to protein purification, but it is hoped that it will also be of value to established practitioners who may find here techniques that they have not tried, but which might well be most applicable in their work. With the exception mainly of the first and last chapters, the format of the contributions to the present book conform to the established format of the Methods in Molecular Biology series.

The introduction of high-performance liquid chromatography (HPLC) to the analysis of peptides and proteins some 25 years ago revolutionized the biological sciences by enabling the rapid and sensitive analysis of peptide and protein structure through the exquisite speed, sensitivity, and resolution that can be easily obtained. Today, HPLC in its various modes has become the pivotal technique in the characterization of peptides and proteins and currently plays a critical role in both our understanding of biological processes and in the development of peptide- and protein-based pharmaceuticals. The number of applications of HPLC in peptide and protein purification continues to expand at an extremely rapid rate. Solid-phase peptide synthesis and recombinant DNA techniques have allowed the production of large quantities of peptides and proteins that need to be highly purified. HPLC techniques are also used extensively in the isolation and characterization of novel proteins that will become increasingly important in the postgenomic age. The design of multidimensional purification schemes to achieve high levels of product purity further demonstrates the power of HPLC techniques not only in the characterization of cellular events, but also in the production of pepti- and protein-based therapeutics. HPLC continues to be at the heart of the analytical techniques with which scientists in both academia and in industry must arm themselves to be able to fully characterize the identity, purity, and potency of peptides and proteins.

Short Protocols in Protein Science provides condensed descriptions of more than 500 protocols compiled from Current Protocols in Protein Science. Drawing from both the original "core" manual as well as the quarterly update service, this compendium includes all step-by-step descriptions of the principal methods covered in Current Protocols in Protein Science.

With the recent completion of the sequencing of the human genome, it is widely anticipated that the number of potential new protein drugs and targets will escalate at an even greater rate than that observed in recent years. However, identification of a potential target is only part of the process in developing these new next generation protein-based “drugs” that are increasingly being used to treat human disease. Once a potential protein drug has been identified, the next rate-limiting step on the road to development is the production of sufficient authentic material for testing, charact- ization, clinical trials, and so on. If a protein drug does actually make it through this lengthy and costly process, methodology that allows the production of the protein on a scale large enough to meet demand must be implemented. Furthermore, large-scale production must not compromise the authenticity of the final product. It is also nec- sary to have robust methods for the purification, characterization, viral inactivation and continued testing of the authenticity of the final protein product and to be able to formulate it in a manner that retains both its biological activity and lends itself to easy administration. Therapeutic Proteins: Methods and Protocols covers all aspects of protein drug production downstream of the discovery stage. This volume contains contributions from leaders in the field of therapeutic protein expression, purification, characterization, f- mulation, and viral inactivation.

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.

This volume covers an array of techniques available for studying peptide-protein docking and design. The book is divided into four sections: peptide binding site prediction; peptide-protein docking; prediction and design of peptide binding specificity; and the design of inhibitory peptides. The chapters in Modeling Peptide-Protein Interactions: Methods and Protocols cover topics such as the usage of ACCLUSTER and PeptiMap for peptide binding site prediction; AnchorDock and ATTRACT for blind, flexible docking of peptides to proteins; flexible peptide docking using HADDOCK and FlexPepDock; identifying loop-mediated protein-protein interactions using LoopFinder; and protein-peptide interaction design using PinaColada. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary details for successful application of the different approaches and step-by-step, readily reproducible protocols, as well as tips on troubleshooting and avoiding known pitfalls. Cutting-edge and thorough, Modeling Peptide-Protein Interactions: Methods and Protocols provides a diverse and unified overview of this rapidly advancing field of major interest and applicability.