Affinity Chromatography (3^rd Ed., 3rd ed. 2015) Methods and Protocols Methods in Molecular Biology Series, Vol. 1286

The aim of this edition is to introduce the beginner to the basics of affinity chromatography and provide practical knowledge for the development of affinity separation protocols. Affinity Chromatography: Methods and Protocols, Third Edition guides readers through new state of the art protocols, molecular modelling, and the study of ligand-target interactions. Written in the successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and notes on troubleshooting and avoiding known pitfalls.

Authoritative and easily accessible, Affinity Chromatography: Methods and Protocols, Third Edition is designed as a useful resource for those interested in the rapid and quantitative isolation of biomolecules with high purity.

1.      Affinity Chromatography: A Historical

David S. Hage and Ryan Matsuda

Part I: General Modes and Ligands of Affinity Chromatography

2.      Protein Purification by Aminosquarylium Cyanine-Affinity Chromatography

Vânia C. Graça, Fani Sousa, Paulo F. Santos and Paulo S. Almeida

3.      One Step Purification of Phosphinothricin Acetyltransferase using Reactive Dye Affinity Chromatography 

Cunxi Wang, Thomas C. Lee, Kathleen S. Crowley, and Erin Bell

4.      Antibody Purification from Human Plasma by Metal-Chelated Affinity Membranes

Handan Yavuz, Nilay Bereli, Fatma Yılmaz, Canan Armutcu and Adil Denizli

5.      Specific Recognition of Supercoiled Plasmid DNA by Affinity Chromatography using a Synthetic Aromatic Ligand

Catarina Caramelo-Nunes and Cândida T. Tomaz

6.      Cell Affinity Separations on Microfluidic Devices

Yan Gao, Wenjie Li, Ye Zhang and Dimitri Pappas

7.      Aptamer-modified :Magnetic Beads in Affinity Separation of Proteins

Guohong Zhu and Johanna-Gabriela Walter

8.      The Strep-tag System for One-step Purification and High Affinity Detection or Capturing of Proteins 

Thomas Schmidt and Arne Skerra

9.      Robotic High-throughput Purification of Affinity-tagged Recombinant Proteins

Simone C. Wiesler and Robert O. J. Weinzierl

Part II: Matrices in Affinity Chromatography

10.  Macroporous Silica Particles Derivatized for Enhanced Lectin Affinity Enrichment of Glycoproteins

Ben Mann

11.  Immobilized Magnetic Beads based Multi-Target Affinity Selection Coupled with HPLC-MS for Screening Active Compounds from Traditional Chinese Medicine and Natural Products

Yaqi Chen, Zhui Chen and Yi Wang

12.  Mixed-bed Affinity Chromatography: Principles And Methods

Egisto Boschetti and Pier Giorgio Righetti

13.  Preparation and characterization of fluorophenylboronic acid-functionalized Affinity Monolithic Columns for the Selective Enrichment of Cis-diol-containing Biomolecules

Qianjin Li and Zhen Liu

Part III: Macroporous Polymers as Matrix in Affinity Chromatography

14.  Introduction to Macroporous Cryogels

Senta Reichelt

15.  Cryogels with Affinity Ligands as Tools in Protein Purification

Solmaz Hajizadeh and Bo Mattiasson

16.  Direct Capture of His6-tagged Proteins Using Megaporous Cryogels Developed for Metal-ion Affinity Chromatography

Naveen Kumar Singh, Roy N. DSouza, Noor Shad Bibi and Marcelo Fernández-Lahore

17.  Ni(II) Chelated IDA Functionalized Poly(HEMA-GMA) Cryogels for Urease Adsorption

Murat UYGUN

18.  A Novel Chromatographic Media: Histidine Containing Composite Cryogels for HIgG Separation from Human Serum

Gözde Baydemir and Mehmet Odabaşı

19.  Molecularly Imprinted Cryogels for Human Serum Albumin Depletion

Muge Andac, Igor Yu Galaev, Handan Yavuz and Adil Denizli

20.  Interpenetrating Polymer Network Composite Cryogels with Tailored Porous Morphology and Sorption Properties

Ecaterina Stela Dragan and Maria Valentina Dinu

Part IV: Spectroscopy, Binding Studies and Molecular Modelling

21.  Analysis of Drug-Protein Interactions by High-Performance Affinity Chromatography: Interactions of Sulfonylurea Drugs with Normal and Glycated Human Serum Albumin

Ryan Matsuda, Jeanethe Anguizola, Krina S. Hoy and David S. Hage

22.  Accurate protein-peptide Titration Experiments by Nuclear Magnetic Resonance using Low-volume Samples

Christian Köhler, Raphaël Recht, Marc Quinternet, Frederic de Lamotte, Marc-André Delsuc, and Bruno Kieffer

23.  Characterization of the Binding Strengths between Boronic Acids and Cis-Diol-containing Biomolecules by Affinity Capillary Electrophoresis

Chenchen Lü and Zhen Liu

24.  Determination of the Kinetic Rate Constant of Cyclodextrin Supramolecular Systems by High Performance Affinity Chromatography

Jiwen Zhang, Haiyan Li, Lixin Sun, and Caifen Wang

25.  Molecular Modeling of the Affinity Chromatography of Monoclonal Antibodies

Matteo Paloni and Carlo Cavallotti

Includes cutting-edge methods and protocols

Provides step-by-step details essential for reproducible results

Contains key notes and implementation advice from the experts