Protein Sequencing Protocols (Methods in Molecular Biology, 211)

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in the intervening period, and the content of this book has similarly changed, the content of some chapters having been superceded and replaced by other approaches.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.

Bryan John Smith updates his much-acclaimed first edition of Protein Sequencing Protocols with new and refreshed techniques for determining the sequence of proteins and peptides. This edition includes not only novel approaches to the validation of quality assurance methods, reflecting the current importance of biopharmaceuticals, but also offers a guide to the analysis of protein sequence information via the powerful new tools of bioinformatics. Each readily reproducible protocol contains detailed step-by-step instructions to ensure successful experimental results, along with notes on troubleshooting and how to avoid pitfalls. Valuable appendices summarize changes in mass arising from protein modification, as well as consensus sequences for protein modification.
Comprehensive and up-to-date, Protein Sequencing Protocols, Second Edition, provides both novice and expert investigators alike a ready source of easy-to-follow protocols that simplify choosing the most appropriate method for protein sequence determination and thus help assure robust results.