Glycosylation Engineering of Biopharmaceuticals

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 Biology(TM) 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.

Engineering of Therapeutic and Diagnostic O-Glycans on Recombinant Mucin-Type Immunoglobulin Fusion Proteins Expressed in CHO Cells. - Engineering a Human-Like Glycosylation to Produce Therapeutic Glycoproteins Based on 6-Linked Sialylation in CHO Cells. - Glycoengineered Pichia-Based Expression of Monoclonal Antibodies. - N-Glycosylation Humanization for Production of Therapeutic Recombinant Glycoproteins in Saccharomyces cerevisiae. - Engineering the Baculovirus Genome to Produce Galactosylated Antibodies in Lepidopteran Cells. - Nano LC Chips MS/MS for the Characterization of N-Glycopeptides Generated from Trypsin Digestion of a Monoclonal Antibody. - Cetuximab Fab and Fc N-Glycan Fast Characterization Using IdeS Digestion and Liquid Chromatography Coupled to Electrospray Ionization Mass Spectrometry. - Therapeutic Antibody Glycosylation Analysis: A Contract Research Organization Perspective in the Frame of Batch Release or Comparability Support. - Mass Spectrometric Analysis of O-Linked Oligosaccharides from Various Recombinant Expression Systems. - Assessing Fc Glycan Heterogeneity of Therapeutic Recombinant Monoclonal Antibodies Using NP-HPLC. - Application of Capillary Electrophoresis in Glycoprotein Analysis. - Characterization of Glycoprotein Biopharmaceutical Products by Caliper LC90 CE-SDS Gel Technology. - Hydrophobic Interaction Chromatography to Analyze Glycoproteins. - Lectin Glycoprofiling of Recombinant Therapeutic Interleukin-7. - Analysis of Monoclonal Antibodies by Sedimentation Velocity Analytical Ultracentrifugation. - Noncovalent Mass Spectrometry for the Characterization of Antibody/Antigen Complexes. - Conformational Analysis of Recombinant Monoclonal Antibodies with Hydrogen/Deuterium Exchange Mass Spectrometry. - Epitope Mapping of Antibodies by Mass Spectroscopy: A Case Study. - Evaluation of Antibody-Dependent Cell Cytotoxicity Using Lactate Deshydrogenase (LDH) Measurement. - Evaluation of Complement Dependent Cytotoxicity Using ATP Measurement and C1q / C4b Binding. - Capture of the Human IgG1 Antibodies by Protein A for the Kinetic Study of h-IgG / FcgR Interaction Using SPR-Based Biosensor Technology. - Mass Spectrometry Protocol for the Absolute Quantification of a Monoclonal Antibody in Serum with Immunopurification.