Affinity Chromatography Methods and Protocols

This volume covers various aspects of affinity chromatography along with examples of its applications. Chapters guide readers through methodologies to purify a diverse array of molecular targets such as antibodies, extracellular vesicles, recombinant proteins, biomarkers, metabolites, plant organelles, nucleic acids, ligand identification and protocols on building affinity matrix. Written in the format of the highly successful Methods in Molecular Biology series, each chapter includes an introduction to the topic, lists necessary materials and reagents, includes tips on troubleshooting and known pitfalls, and step-by-step, readily reproducible protocols.

Antibody Purification Using Affinity Chromatography.- TIM4-affinity Methods Targeting Phosphatidylserine for Isolation or Detection of Extracellular Vesicles.- Application of Affitins for Affinity Purification of Proteins.- Identification of Clusterin as a major ABri- and ADan-binding protein using affinity Chromatography.- Application of a Novel CL7/Im7 Affinity System in Purification of Complex and Pharmaceutical Proteins.- Positive and Negative Affinity Chromatography to Purify Norovirus P-Domain Protein.- Robotic Affinity Purification of Soluble and Insoluble Recombinant Glutathione-S-Transferase Fusion Proteins.- Application of Immunoaffinity-Mass Spectrometry (IA-MS) for Protein Biomarker Quantification.- Cellular Membrane Affinity Chromatography for the Identification of Bioactive Molecules from Complex Natural Mixtures.- Rapid Affinity Purification of Tagged Plant Mitochondria (Mito-AP).- Arginine-affinity Chromatographyfor Nucleic Acids (DNA and RNA) Isolation.- Purification and Analysis of Nucleotides and Nucleosides from Plants.- Ligand Selection for Affinity Chromatography Using Phage Display.- Selection and Application of Aptamer Affinity for Protein Purification.- Entrapment of Proteins within Columns for High-Performance Affinity Chromatography.- Preparation of Affinity Chromatography Monolith in Miniaturized Format and Application for Protein Separation.- Ligand Immobilization Methods for Affinity Chromatography.- Preparation of Molecularly Imprinted poly(N-isopropylacrylamide) Thermosensitive Based Cryogels.- Preparation of Staphylococcal Protein a Imprinted Supermacroporous Cryogel Beads.