Glyco-Engineering, Softcover reprint of the original 1st ed. 2015 Methods and Protocols Methods in Molecular Biology Series, Vol. 1321

Conceived with the intention of providing an array of strategies and technologies currently in use for glyco-engineering distinct living organisms, this book contains a wide range of methods being developed to control the composition of carbohydrates and the properties of proteins through manipulations on the production host rather than in the protein itself. The first five sections deal with host-specific glyco-engineering and contain chapters that provide protocols for modifications of the glycosylation pathway in bacteria, yeast, insect, plants and mammalian cells, while the last two sections explore alternative approaches to host glyco-engineering and selected protocols for the analysis of the N-glycans and glyco-profiling by mass spectrometry. Written for the highly successful Methods in Molecular Biology series, 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 extensive, Glyco-Engineering: Methods and Protocols offers vast options to help researchers to choose the expression system and approach that best suits their intended protein research or applications.

Part I: Glyco-Engineering in Bacteria

1. Current Approaches to Engineering N-Linked Protein Glycosylation in Bacteria

            Andreas Naegeli and Markus Aebi

2. Inverse Metabolic Engineering for Enhanced Glycoprotein Production in Escherichia coli

            Stephen R.P. Jaffé, Benjamin Strutton, Jagroop Pandhal, and Phillip C. Wright

3. GlycoSNAP: A High-Throughput Screening Methodology for Engineering Designer Glycosylation Enzymes

            Anne A. Ollis, Yi Chai, and Matthew P. DeLisa

4. Production of Glycoproteins with Asparagine-Linked N-Acetylglucosamine in Escherichia coli

            Flavio Schwarz and Markus Aebi

5. Glyco-Engineering O-Antigene-Based Vaccines and Diagnostics in E. coli

            Nicolas F. Vozza and Mario F. Feldman

Part II: Glyco-Engineering in Yeast

6. Progress in Yeast Glycosylation Engineering

            Stephen R. Hamilton and Dongxing Zha

7. Protein Production with a Pichia pastoris OCH1 Knock-Out Strain in Fed-Batch Mode

            Christoph Gmeiner and Oliver Spadiut

8. Engineering the Pichia pastoris N-Glycosylation Pathway Using the GlycoSwitch Technology

            Bram Laukens, Charlot De Wachter, and Nico Callewaert

9. Development of a Valuable Yeast Strain Using a Novel Mutagenesis Technique for the Effective Production of Therapeutic Glycoproteins

            Hiroko Abe and Ken-ichi Nakayama

Part III: Glyco-Engineering in Insects

10. An Overview and History of Glyco-Engineering in Insect Expression Systems

            Christoph Geisler, Hideaki Mabashi-Asazuma, and Donald L. Jarvis

11. SweetBac: Applying

            Dieter Palmberger and Dubravko Rendic

12. Engineering Sialic Acid Synthesis Ability in Insect Cells

            Karthik Viswanathan, Someet Narang, and Michael J. Betenbaugh

13. Engineering N-Glycosylation Pathway in Insect Cells: Suppression of β-N-Acetylglucosaminidase and Expression of β-1,4-Galactosyltransferase

            Yeon Kyu Kim and Hyung Joon Cha

Part IV: Glyco-Engineering in Plants

14. N-Glyco-Engineering in Plants: Update on Strategies and Major Achievements

            Herta Steinkellner and Alexandra Castilho

15. Gene Targeting for Precision Glyco-Engineering: Production of Biopharmaceuticals Devoid of Plant-Typical Glycosylation in Moss Bioreactors

            Eva L. Decker, Gertrud Wiedemann, and Ralf Reski

16. Im“plant”ing of Mammalian Glycosyltransferase Gene into Plant Suspension-Cultured Cells Using Agrobacterium-Mediated Transformation

            Hiroyuki Kajiura and Kazuhito Fujiyama

17. Transient Glyco-Engineering of N. benthamianaAiming at the Synthesis of Multi-Antennary Sialylated Proteins

            Andreas Loos and Alexandra Castilho

18. Subcellular Targeting of Proteins Involved in Modification of plant N- and O-Glycosylation

            Martina Dicker, Jennifer Schoberer, Ulrike Vavra, and Richard Strasser

19. Assembly of Multigene Constructs Using Golden Gate Cloning

            Sylvestre Marillonnet and Stefan Werner

Part V: Glyco-Engineering in Mammalians

20. Strategies for Engineering Protein N-Glycosylation Pathway in Mammalian Cells

            Qiong Wang, Mark Stuczynski, Yuan Gao, and

21. Glycan Remodelling with Processing Inhibitors and Lectin-Resistant Eukaryotic Cells

            Veronica T. Chang, Robert A. Spooner, Max Crispin, and Simon J. Davis

22. Production of Highly Sialylated Recombinant Glycoproteins Using Ricinus communis Agglutinin-I-Resistant CHO Glycosylation Mutants

            John S.Y. Goh, Kah Fai Chan, and Zhiwei Song

23. Metabolic Glyco-Engineering in Eukaryotic Cells and Selected Applications

            Friedrich Piller, Aline Mongis, and Véronique Piller

24. Evaluation of Quenching and Extraction Methods for Nucleotide/Nucleotide Sugar Analysis

            Katrin Braasch, Carina Villacrés Barragán, and Michael Butler

Part VI: In Vitro Glyco-Engineering of Recombinant Proteins

25. Chemoenzymatic Glyco-Engineering of Monoclonal Antibodies

            John P. Giddens and Lai-Xi Wang

26. Chemical Polysialylation of Recombinant Human Proteins

            Ivan V. Smirnov, Ivan I. Vorobiev, Alexey A. Belogurov, Dmitry D. Genkin, Sergey M. Deyev, and Alexander G. Gabibov

Part VII: Glycan Analysis

27. Site Specific Glycosylation Profiling Using Liquid Chromatography-Tandem Mass Spectrometry (LC-MS)

            Clemens Gruber and Friedrich Altmann

28. Mass Spectrometric Analysis of Oligo- and Polysialic Acids

            Christina E. Galuska, Kai Maass, and Sebastian P. Galuska

29. Isomer-Specific Analysis of Released N-Glycans by LC-ESI MS/MS with Porous Graphitized Carbon

            Daniel Kolarich, Markus Windwarder, Kathirvel Alagesan, and Friedrich Altmann

Features protocols in glyco-engineering in bacteria, yeast, insects, plants and mammalian cells

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts