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This detailed book covers methods for studying, producing, and analyzing therapeutic antibodies, measuring their concentration, developing neutralizing antibodies for them, and for predicting and monitoring their therapeutic efficacy and clinical effects. These biologics are the fastest growing pharmaceutical drug group and have had tremendous clinical and scientific impact in cancer, autoimmune diseases, infectious diseases, and other immune-related diseases, making the content of this volume essential. 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 methods, and tips on troubleshooting and avoiding known pitfalls.
Authoritative and practical, Therapeutic Antibodies: Methods and Protocols serves as an ideal guide for researchers working with the production of, research on, and development of therapeutic antibodies as well as for clinicians using therapeutic antibodies in daily work with patients.
List of contents
Therapeutic Antibodies: An Overview.- Treatment of Autoimmune Diseases with Therapeutic Antibodies: Lessons Learned from PID Patients Allow for Stratification of the Infection Risk.- B-Cell Analysis for Monitoring Patients Undergoing B-Cell Depletion for the Treatment of Autoimmune Diseases.- Assessment of Therapeutic Antibody Developability by Combinations of In Vitro and In Silico Methods.- The Therapeutic Antibody Profiler for Computational Developability Assessment.- Short Read-Length Next-Generation DNA Sequencing of Antibody CDR Combinations from Phage Selection Outputs.- Large Scale Transient Production in ExpiCHO-STM with Enhanced N-Galactosylation-Sialylation and PEI-Based Transfection.- Production of Therapeutic Single-Chain Variable Fragments (ScFv) in Pichia pastoris.- Purification of Therapeutic Antibodies by Protein A Affinity Chromatography.- Ion Exchange Chromatographic Methods for Purification of Therapeutic Antibodies.- Structure-Indicated LC-MS/MS Bioanalysis of Therapeutic Antibodies.- Use of PASEF for Accelerated Protein Sequence Confirmation and De Novo Sequencing with High Data Quality.- Nano-Microscopy of Therapeutic Antibody Aggregates in Solution.- Measuring Self-Association of Antibody Lead Candidates with Dynamic Light Scattering.- Intact Mass Quantitation of Therapeutic Antibodies for Pharmacokinetic Studies Using Immuno-Purification.- Detection of Residual Host Cell Proteins in Biotherapeutic Drugs by Concatenated 2D LC-MS/MS.- Bioassays for the Evaluation of Target Neutralization and Complement-Dependent Cytotoxicity (CDC) of Therapeutic Antibodies.- Methods for Functional Characterization of FcRn Interactions with Therapeutic Antibodies and Fc-Fusion Proteins.- Method for Measurement of Antibody-Dependent Cellular Phagocytosis.- Bioluminescent Bridging Immunoassay for Anti-Drug Antibody (ADA) Detection.- A Surface Plasmon Resonance-Based Assay for Simultaneous Measurement of Serum Concentrations of Therapeutic Antibodies and Anti-Drug Antibodies.
