Affinity Chromatography, 1st ed. 2022
Methods and Protocols
Methods in Molecular Biology Series, Vol. 2466

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.

Part I: Applications of Affinity Chromatography

1. Antibody Purification Using Affinity Chromatography

Arabelle Cassedyand Richard O'Kennedy

2. TIM4-affinity Methods Targeting Phosphatidylserine for Isolation or Detection of Extracellular Vesicles

Takeshi Yoshidaand Rikinari Hanayama

3. Application of Affitins for Affinity Purification of Proteins

Barbara Mouratou and Frédéric Pecorari

4. Identification of Clusterin as a major ABri- and ADan-binding protein using affinity Chromatography

Agueda Rostagno, Miguel Calero, and Jorge Ghiso

5. Application of a Novel CL7/Im7 Affinity System in Purification of Complex and Pharmaceutical Proteins

Louise T. Chow and Dmitry G. Vassylyev

6. Positive and Negative Affinity Chromatography to Purify Norovirus P-Domain Protein

Wilhelm Salmen

7. Robotic Affinity Purification of Soluble and Insoluble Recombinant Glutathione-S-Transferase Fusion Proteins

Robert O.J. Weinzierl

8. Application of Immunoaffinity-Mass Spectrometry (IA-MS) for Protein Biomarker Quantification

Joe Palandra, Nikolaos Psychogios, and Hendrik Neubert

9. Cellular Membrane Affinity Chromatography for the Identification of Bioactive Molecules from Complex Natural Mixtures

Z. Ceren Arituluk and Lukasz M. Ciesla

Markus Niehaus and Marco Herde

11. Arginine-affinity Chromatography for Nucleic Acids (DNA and RNA) Isolation

Ana Rita Carapito, Joana F. A. Valente,João A. Queiroz, and Fani Sousa

12. Purification and Analysis of Nucleotides and Nucleosides from Plants

Henryk Straube and Marco Herde

Part II: Ligand Selection and Affinity Matrix Preparation for Affinity Chromatography

13. Ligand Selection for Affinity Chromatography Using Phage Display

Krištof Bozovičar, Peter Molek, Barbara Jenko Bizjan, and Tomaž Bratkovič

14. Selection and Application of Aptamer Affinity for Protein Purification

Ana Paula de Jesus Santos, Ágatha Oliveira-Giacomelli, Vanessa Karen de Sá, Isis Cristina do Nascimento, Erika de Simone Molina, and Henning Ulrich

15. Entrapment of Proteins within Columns for High-Performance Affinity Chromatography

Saumen Poddar, Sadia Sharmeen, and David S. Hage

16. Preparation of Affinity Chromatography Monolith in Miniaturized Format and Application for Protein Separation

Kishore K. R. Tetala

17. Ligand Immobilization Methods for Affinity Chromatography

Chandra K. Dixit

18. Preparation of Molecularly Imprinted poly(N-isopropylacrylamide) Thermosensitive Based Cryogels

Işık Perçin, Neslihan Idil, and Adil Denizli

19. Preparation of Staphylococcal Protein a Imprinted Supermacroporous Cryogel Beads

Sevgi Aslıyüce, Bo Mattiasson, and Adil Denizli

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts