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This volume explores protocols and perspectives for the design, production, characterization, functional evaluation, and clinical translation of bispecific antibodies (BsAbs). Beginning with a variety of platforms for generating BsAbs, the book continues with sections covering protocols to evaluate T-cell-dependent bispecific antibodies, modeling and simulation approaches to support clinical development of T-cell-engaging BsAbs, as well as clinical applications. 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 and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Bispecific Antibodies: Methods and Protocols aims to bridge the gap between molecular design and translational success by equipping investigators with the technical know-how and contextual understanding needed to show each method s relevance to therapeutic development.
List of contents
Production of Bispecific Antibody Combining Knobs-into-Holes and Controlled Fab-Arm Exchange Methods.- Multimodal Physicochemical Characterization of Cross-Linking Activities of Biparatopic Antibodies.- Generation of Bispecific Antibodies Using FAST-Ig .- ImmTAC: A Novel Platform of T-Cell Receptor-Based Soluble Bispecifics.- Design and Evaluation of Domain-Rearranged Bispecific Antibody Fragments for Functional Optimization.- Production of Bispecific Antibody Fragments from Fc Fusion.- Enhanced Chemical Method for Bispecific Antibody Production: Leveraging Site-Specific Conjugation and Stable Crosslinker Technology.- Laboratory-Scale Production of Recombinant IgG Antibodies.- Visualization of Immunological Synapse Formation by Intercellular Cross-Linking of T-Cell-Dependent Bispecific Antibodies Using Fluorescence Microscopy.- Measurement of T-Cell Migration by T-Cell-Dependent Bispecific Antibody in Cancer Cells and T-Cell Cross-Linking.- Optimized In Vitro T-Cell Exhaustion Assay for Evaluating T-Cell-Dependent Bispecific Antibodies.- Advanced Evaluation of T-Cell-Dependent Bispecific Antibodies Using Fluorescence Imaging and In Vivo Models.- Fluorescent-Based Analysis of T-Cells and T-Cell-Dependent Bispecific Antibody in a Brain Tumor Mouse Model.- Peripheral Blood Mononuclear Cell (PBMC) Humanized Mice as a Platform for a Preclinical In Vivo Evaluation of T-Cell-Dependent Bispecific Antibodies.- Overview of Quantitative Clinical Pharmacology for T-Cell Engaging Bispecific Antibodies.- Clinical Development of T-Cell Bispecific Antibodies: Review, Challenges, and Prospective Solutions in Solid Tumor.- Clinical Development of Bispecific Antibodies: Review, Advances, and Challenges in Hematological Malignancies.
Summary
This volume explores protocols and perspectives for the design, production, characterization, functional evaluation, and clinical translation of bispecific antibodies (BsAbs). Beginning with a variety of platforms for generating BsAbs, the book continues with sections covering protocols to evaluate T-cell-dependent bispecific antibodies, modeling and simulation approaches to support clinical development of T-cell-engaging BsAbs, as well as clinical applications. 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 and readily reproducible laboratory protocols, and tips for troubleshooting and avoiding known pitfalls.
Authoritative and practical, Bispecific Antibodies: Methods and Protocols aims to bridge the gap between molecular design and translational success by equipping investigators with the technical know-how and contextual understanding needed to show each method’s relevance to therapeutic development.
