This volume explores the latest integrated bioprocesses and technologies used to study the production of the target recombinant protein of therapeutic or diagnostic interest; its isolation, purification, and stabilization; and the bio-interaction and structural analyses. The chapters in this book are organized into four parts. Part One covers production methods of soluble and membrane proteins in prokaryotic and eukaryotic expression systems, such as Lactococcus lactis and Escherichia coli. Part Two describes traditional and novel approaches for recombinant protein purification and stabilization, and buffers and additives. Part Three discusses automated methods in structural biology based on in silico approaches; and Part Four provides examples of advanced protein investigation methodologies to assess structural analysis such as high-throughput protein crystallization and time-resolved serial crystallography. Written in the highly successful Methods in Molecular Biology series format, 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.Cutting-edge and comprehensive, Advanced Methods in Structural Biology is a valuable resource to those in academia (i.e., graduate students and postdoctoral researchers) and 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.
Reihe
Sprache
Verlagsort
Verlagsgruppe
Illustrationen
82
21 s/w Abbildungen, 82 farbige Abbildungen
XV, 467 p. 103 illus., 82 illus. in color.
Dateigröße
ISBN-13
978-1-0716-3147-8 (9781071631478)
DOI
10.1007/978-1-0716-3147-8
Schweitzer Klassifikation
Cloning and Recombinant Protein Expression in Lactococcus lactis.- E6 Tagged Protein Production, Extraction, and Purification from Escherichia coli Lysate.- Mini-Bioreactor Platform for Membrane Proteins Production in Komagataella pastoris.- Recombinant Protein Production Using the Baculovirus Expression Vector System (BEVS).- High-Throughput Production and Optimization of Membrane Proteins after Expression in Mammalian Cells.- Fluorescent Fusion Proteins Expression in Plant Cells.- Functional Expression of the Ectodomain of Plant Receptor Kinases in Plant Suspension Culture.- A Simple Purification Method for Heat Stable Recombinant Low Molecular Weight Proteins and Peptides Via GST-Fusion Products.- eGFP as an All-in-One Tag for Purification of Membrane Proteins.- Effects of Ionic Liquids as Additives on Protein Crystallization.- Screening of Buffers and Additives for Protein Stabilization by Thermal Shift Assay: A Practical Approach.- Biophysical Characterization of Membrane Proteins.- Expression in Saccharomyces cerevisiae and Purification of a Human Phospholipid Flippase.- Preparation of Protein-Enriched Outer Membrane Vesicles from Escherichia Coli for In Situ Structural Biology of Outer Membrane Proteins.- Receptor-Based Virtual Screening of Large Libraries in a Multi-Level in Silico Approach.- Protein-Protein Interaction Studies Using Molecular Dynamics Simulation.- Isolation of Bacterial Extracellular Vesicles and Identification of their Protein Cargo.- Study of Protein Conformational Dynamics Using Hydrogen/Deuterium Exchange Mass Spectrometry.- Protein Interaction Analysis by Surface Plasmon Resonance.- High-Throughput Protein Crystallization in an Integrated Droplet-Based Microfluidic Platform.- Sample Preparation for Time-Resolved Serial Crystallography: Practical Considerations.- Using Small-Angle X-Ray Scattering to Characterize Biological Systems:A General Overview and Practical Tips.- Nuclear Magnetic Resonance-Guided Structural Analysis of Moderate-Affinity Protein Complexes with Intrinsically Disordered Polypeptides.- Protein Structural Analysis by Cryogenic Electron Microscopy.
