Synthetic Antibodies

This detailed volume presents a set of protocols useful for researchers in the field of recombinant immunoglobulin and alternative scaffold engineering, aptamer development, and generation of molecularly imprinted polymers (MIPs). Part I includes methods that deal with amino-acid based synthetic antibodies. Brief protocols about the generation of antibody libraries are detailed, as well as techniques for antibody selection, characterization, and validation. This section is completed by a brief description of a bioinformatics platform that supports antibody engineering during research and development. Part II contains basic procedures about the selection and characterization of aptamer molecules, and Part III describes fundamental processes of MIP generation and application. 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 practical, Synthetic Antibodies: Methods and Protocols is an ideal guide for scientists seeking to propel the vital study of antibody research.

Antibody Mimetics, Peptides, and Peptidomimetics.- Construction of a scFv Library with Synthetic, Non-Combinatorial CDR Diversity.- Enzymatic Assembly for scFv Library Construction.- Directed Evolution of Protein Thermal Stability Using Yeast Surface Display.- Whole Cell Panning with Phage Display.- Generating Conformation and Complex-Specific Synthetic Antibodies.- High-Throughput IgG Conversion of Phage Displayed Fab Antibody Fragments by AmplYFast.- Utilization of Selenocysteine for Site-Specific Antibody Conjugation.- Solubility Characterization andImaging of Intrabodies Using GFP-Fusions.- Antibody Validation by Immunoprecipitation Followed by Mass Spectrometry Analysis.- Novel HPLC-Based Screening Method to Assess Developability of Antibody-Like Molecules.- Glycosylation Profiling of /ß T Cell Receptor Constant Domains Expressed in Mammalian Cells.- A Proximity-Based Assay for Identification of Ligand and Membrane Protein Interaction in Living Cells.- A Biotin Ligase-Based Assay for the Quantification of the Cytosolic Delivery of Therapeutic Proteins.- Data-Driven Antibody Engineering Using Genedata Biologics .- Selection of Aptamers Against Whole Living Cells: From Cell-SELEX to Identification of Biomarkers.- Rapid Selection of RNA Aptamers that Activate Fluorescence of Small Molecules.- An Enzyme-Linked Aptamer Sorbent Assay to Evaluate Aptamer Binding.- Incorporating Aptamers in the Multiple Analyte Profiling Assays (xMAP): Detection of C-Reactive Protein.- Transferring the Selectivity of a Natural Antibody into a Molecularly Imprinted Polymer.- Preparation of Molecularly Imprinted Microspheres by Precipitation Polymerization.- Generation of Janus Molecularly Imprinted Polymer Particles.- Surface Engineering of Nanoparticles to Create Synthetic Antibodies.- H5N1 Virus Plastic Antibody Based on Molecularly Imprinted Polymers.- Replacement of Antibodies in Pseudo-ELISAs: Molecularly Imprinted Nanoparticles for Vancomycin Detection.- Cell and Tissue Imaging with Molecularly Imprinted Polymers.