Glycoprotein Methods and Protocols

Glycoprotein Methods and Protocols
Author: Anthony P. Corfield
Publisher: Springer Science & Business Media
Total Pages: 491
Release: 2007-10-26
Genre: Science
ISBN: 1592590489

The mucins (mucus glycoproteins) have long been a complex corner of glycoprotein biology. While dramatic advances in the separation, structural an- ysis, biosynthesis, and degradation have marked the progress in general glycop- tein understanding, the mucins have lagged behind. The reasons for this lack of progress have always been clear and are only now being resolved. The mucins are very large molecules; they are difficult to separate from other molecules present in mucosal secretions or membranes; they are often degraded owing to natural protective functions or to isolation methodology and their peptide and oligos- charide structures are varied and complex. Understanding these molecules has demanded progress in several major areas. Isolation techniques that protect the intact mucins and allow dissociation from other adsorbed but discrete molecules needed to be developed and accepted by all researchers in the field. Improved methods for the study of very large molecules with regard to their aggregation and polymerization were also needed. Structural analysis of the peptide domains and the multitude of oligosaccharide chains was required for smaller sample sizes, for multiple samples, and in shorter time. In view of these problems it is perhaps not surprising that the mucins have remained a dilemma, of obvious biological importance and interest, but very difficult to analyze.

Mass Spectrometry of Glycoproteins

Mass Spectrometry of Glycoproteins
Author: Arnaud Delobel
Publisher: Humana
Total Pages: 403
Release: 2021-07-04
Genre: Science
ISBN: 9781071612408

This volume presents methods used for the analysis of glycoproteins at different levels—intact, subunit, glycopeptide, and monosaccharide--, and discusses and solves most analytical challenges that a scientist working on glycoproteins may come across. The chapters in this book cover topics such as the role of glycosylation on the properties of therapeutic glycoproteins; different analytical methods to characterize glycosylation, from the intact proteins to the glycan level, for both N-linked and O-linked glycoproteins; mass spectrometry imaging methodology for glycosylation analysis in tissues; approaches to characterizing glycosylation on cultured cells; and the use of cloud computing to deploy mass spectrometry data analysis. 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 thorough, Mass Spectrometry of Glycoproteins: Methods and Protocols is a valuable resource for scientists interested in learning more about this developing field.

Glycosylation Engineering of Biopharmaceuticals

Glycosylation Engineering of Biopharmaceuticals
Author: Alan Beck
Publisher: Humana Press
Total Pages: 0
Release: 2013-03-09
Genre: Medical
ISBN: 9781627033268

Glyco-engineering is being developed as a method to control the composition of carbohydrates and to enhance the pharmacological properties of monoclonal antibodies (mAbs) and other proteins. In Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols, experts in the field provide readers with production and characterization protocols of glycoproteins and glyco-engineered biopharmaceuticals with a focus on mAbs. The volume is divided in four complementary parts dealing with glyco-engineering of therapeutic proteins, glycoanalytics, glycoprotein complexes characterization, and PK/PD assays for therapeutic antibodies. Written in the highly successful Methods in Molecular BiologyTM 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. Authoritative and cutting-edge, Glycosylation Engineering of Biopharmaceuticals: Methods and Protocols serves as an ideal guide for scientists striving to push forward the exciting field of engineered biopharmaceuticals.

Pichia Protocols

Pichia Protocols
Author: James M Cregg
Publisher: Springer Science & Business Media
Total Pages: 553
Release: 2007-08-08
Genre: Science
ISBN: 1588294293

This book focuses on recent developments of Pichia pastoris as a recombinant protein production system. Highlighted topics include a discussion on the use of fermentors to grow Pichia pastoris, information on the O- and N-linked glycosylation, methods for labeling Pichia pastoris expressed proteins for structural studies, and the introduction of mutations in Pichia pastoris genes by the methods of restriction enzyme-mediated integration (REMI). Each chapter presents cutting-edge and cornerstone protocols for utilizing P. pastoris as a model recomibinant protein production system. This volume fully updates and expands upon the first edition.

Glycomics

Glycomics
Author: Nicolle H. Packer
Publisher: Humana Press
Total Pages: 390
Release: 2009-07-16
Genre: Science
ISBN: 9781607612407

Due to the significant contributions of carbohydrates to the functional diversity of the cell, the challenging study of the glycome has expanded beyond the research of carbohydrate experts and into the wider scope of the life sciences. To aid all scientists now delving into this vital subject area, Glycomics: Methods and Protocols collects a compendium of detailed laboratory protocols reflecting the increasing availability of sample preparation, chromatographic, electrophoretic, mass spectrometric, and bioinformatic tools specifically designed for the analysis of glycosylation. Leading researchers in the field address subjects such as glycoprotein and proteoglycan analysis, glycosylation structure determination, as well as various approaches to investigate the interaction between glycans and a variety of carbohydrate-recognizing proteins in order to aid exploration into the functional significance of the oligosaccharides. Written in the highly successful Methods in Molecular BiologyTM series format, the chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Glycomics: Methods and Protocols serves as a valuable guide for experimenters facing the challenges of glycan analysis in hope of providing further insights into the biology of cell-cell communication and interaction.

Essentials of Glycobiology

Essentials of Glycobiology
Author: Ajit Varki
Publisher: CSHL Press
Total Pages: 694
Release: 1999
Genre: Medical
ISBN: 9780879696818

Sugar chains (glycans) are often attached to proteins and lipids and have multiple roles in the organization and function of all organisms. "Essentials of Glycobiology" describes their biogenesis and function and offers a useful gateway to the understanding of glycans.

Monoclonal Antibodies

Monoclonal Antibodies
Author: Maher Albitar
Publisher: Springer Science & Business Media
Total Pages: 472
Release: 2008-02-02
Genre: Medical
ISBN: 1597453234

This book examines a collection of state-of-the-art methods that employ monoclonal antibodies in a clinical setting. The chapters offer in-depth description for generating mouse and recombinant humanized antibodies, and a comprehensive review of how antibodies are being used in bead-based methods for measuring proteins. This field will continue to expand and provide new and innovative techniques in the laboratory and as a basis that complements targeted therapy.

Insoluble Proteins

Insoluble Proteins
Author: Elena García-Fruitós
Publisher: Humana Press
Total Pages: 0
Release: 2014-12-02
Genre: Science
ISBN: 9781493922048

With insolubility proving to be one of the most crippling bottlenecks in the protein production and purification process, this volume serves to aid researchers working in the recombinant protein production field by describing a wide number of protocols and examples. Insoluble Proteins: Methods and Protocols includes chapters that describe not only the recombinant protein production in different expression systems but also different purification and characterization methods to finally obtain these difficult-to-obtain proteins. Beginning with protein production methods using both prokaryotic and eukaryotic expression systems, the book continues with purification protocols using insoluble proteins, the characterization of insoluble proteins, as well as a general overview of interesting applications of insoluble proteins. 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. Comprehensive and practical, Insoluble Proteins: Methods and Protocols aims to provide the scientific community with detailed and reliable state-of-the-art protocols that are used in order to successfully produce and purify recombinant proteins prone to aggregate.

Lectins

Lectins
Author: Jun Hirabayashi
Publisher: Humana Press
Total Pages: 613
Release: 2014-08-13
Genre: Science
ISBN: 9781493912919

Lectins: Methods and Protocols summarizes classic lectin technologies and advanced techniques with high throughputs and sensitivities. Chapters include methods and techniques for serial lectin-affinity chromatography procedure, lectin-probed western blot and histochemical analyses, quantitative interaction analyses based on equilibrium dialysis, isothermal calorimetry, surface plasmon resonance, techniques for elucidating functions of endogenous animal lectins, advanced methods on “engineer” novel lectins by evolutionary concepts, and special equipment. 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 key tips on troubleshooting and avoiding known pitfalls. Authoritative and practical, Lectins: Methods and Protocols seeks to aid not only lectin specialists but also non-experts including both young scientists.

Clinical Proteomics

Clinical Proteomics
Author: Antonia Vlahou
Publisher: Humana Press
Total Pages: 0
Release: 2014-11-11
Genre: Science
ISBN: 9781493918713

This second edition expands upon the previous edition with current, detailed developments in the field and brings together a multi-disciplinary team of leading researchers to provide their latest protocols for clinical proteomics analysis. 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. Authoritative and Practical, Clinical Proteomics: Methods and Protocols, Second Edition presents approaches that will serve as a reliable guide to researchers, including clinicians, chemists, molecular biologists, bioinformaticians and computational, biologists, and investigators working on biomarker development.