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This detailed volume explores the production and purification of glycoproteins for therapeutic use or basic research, as well as the necessary in-depth understanding of cellular and acellular systems available for these purposes, their advantages and disadvantages, and considerations for choosing the most appropriate system for the desired application. Beginning with chapters on viral-encoded glycoproteins and those found in other pathogens, the book continues by examining the production of mammalian glycoproteins, the analysis of glycoprotein content, cell-free synthesis of glycoproteins and considerations for production and purification of glycoproteins. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to the respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Recombinant Glycoproteins: Methods and Protocols provides a wide range of guidelines for studying these vitally important proteins.
Includes cutting-edge techniques Provides step-by-step detail essential for reproducible results Contains key implementation advice from the experts
Contenu
Production and Purification of Hantavirus Glycoproteins in Drosophila melanogaster S2 Cells.- Production and Purification of Filovirus Glycoproteins.- Modification of N-Linked Glycan Sites in Viral Glycoproteins.- Production of Influenza Virus Glycoproteins Using Insect Cells.- SARS-CoV-2 S-Protein Ace2 Binding Analysis Using Surface Plasmon Resonance.- A Biosensor Assay Based on Coiled-Coil-Mediated Human ACE2 Receptor Capture for the Analysis of Its Interactions with the SARS-CoV-2 Receptor Binding Domain.- Production and Purification of Plasmodium Circumsporozoite Protein in Lactococcus lactis .- Analysis of Caenorhabditis Protein Glycosylation.- Use of Reductive Amination to Produce Capsular Polysaccharide-Based Glycoconjugates.- Overexpression and Purification of Mitogenic and Metabolic Fibroblast Growth Factors.- Production and Purification of Antibodies in Chinese Hamster Ovary Cells.- Mammalian Antigen Display for Pandemic Countermeasures.- Analysisof Native and Permethylated N-Glycan Isomers Using MGC-LC-MS Techniques.- Targeted Glycoproteomics Analysis Using MRM/PRM Approaches.- Targeted Analysis of Permethylated N -Glycans Using MRM/PRM Approaches.- Hydrophilic Interaction Liquid Chromatography (HILIC) Enrichment of Glycopeptides Using PolyHYDROXYETHYL A.- O -Glycoproteomics Sample Preparation and Analysis Using NanoHPLC and Tandem MS.- Solubilization of Oligomeric Cell-Free Synthesized Proteins Using SMA Copolymers.- Cell-Free Systems for the Production of Glycoproteins.- Considerations for Glycoprotein Production.