Advanced Methods in Structural Biology

This volume explores the latest integrated bioprocesses and technologies used to study the production of recombinant proteins, their isolation, purification, and stabilization, and their bio-interactions and structural analyses. It is written in the highly successful Methods in Molecular Biology series format and is a valuable resource for those in academia and the pharmaceutical industry.

Format: Hardback
Length: 467 pages
Publication date: 25 April 2023
Publisher: Springer-Verlag New York Inc.

This comprehensive volume delves into the latest integrated bioprocesses and technologies employed to investigate the production of therapeutic or diagnostic recombinant proteins. It encompasses the isolation, purification, and stabilization of these proteins, as well as the exploration of their bio-interactions and structural analyses. The book is organized into four parts, each covering a distinct aspect of protein production and characterization.

Part One explores the methods used to produce soluble and membrane proteins in prokaryotic and eukaryotic expression systems, such as Lactococcus lactis and Escherichia coli. It discusses the techniques employed to optimize protein expression, including the use of expression vectors, host strains, and growth conditions. The chapter also highlights the importance of protein purification and stabilization, emphasizing the use of various chromatographic methods and additives to enhance protein yield and stability.

Part Two delves into traditional and novel approaches for recombinant protein purification and stabilization. It discusses the use of affinity chromatography, ion exchange chromatography, and size exclusion chromatography, as well as the application of various buffers and additives to improve protein solubility and stability. The chapter also highlights the importance of buffer selection and pH adjustment to maintain protein activity and prevent denaturation.

Part Three explores automated methods in structural biology based on in silico approaches. It discusses the use of computational tools and algorithms to predict protein structures, as well as the techniques employed to obtain experimental structures through methods such as X-ray crystallography and NMR spectroscopy. The chapter also highlights the importance of structural analysis in understanding protein function and interactions, and the use of advanced techniques such as high-throughput protein crystallization and time-resolved serial crystallography to accelerate the structural characterization process.

Part Four provides examples of advanced protein investigation methodologies to assess structural analysis, such as high-throughput protein crystallization and time-resolved serial crystallography. It discusses the experimental setup, data collection, and analysis techniques employed in these methods, as well as the challenges and limitations associated with each approach. The chapter also highlights the applications of structural analysis in drug discovery, protein engineering, and disease modeling.

Written in the highly successful Methods in Molecular Biology series format, the chapters in this book 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. The book is designed to be a valuable resource for those in academia, including graduate students and postdoctoral researchers, as well as researchers in the pharmaceutical industry who wish to learn more about this developing field.

Chapter 5 is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Weight: 1099g
Dimension: 254 x 178 (mm)
ISBN-13: 9781071631461
Edition number: 1st ed. 2023