Glycoprotein Methods And Protocols

Glycoprotein Methods and Protocols

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.

Recombinant Glycoproteins

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. Chapter “Solubilization of Oligomeric Cell-Free Synthesized Proteins Using SMA Copolymers” is available open access under a Creative Commons Attribution 4.0 International License via link.springer.com.

Methods in Molecular Biology: Glycoprotein methods and protocols : the mucins