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Published on: January 8, 2014
Purification of antibodies using affinity chromatography
Elaine Darcy1, Paul Leonard, Jenny Fitzgerald
1School of Biotechnology, Dublin City University, Dublin, Ireland.
Methods in Molecular Biology (Clifton, N.J.)
|October 28, 2010
Summary
Affinity chromatography purifies target molecules using specific binding pairs. This guide details immuno-affinity chromatography for antibody purification using proteins A and G.
Area of Science:
- Biochemistry
- Molecular Biology
- Chromatography
Background:
- Affinity chromatography isolates target analytes from complex mixtures.
- It relies on specific interactions between binding affinity pairs.
- Immuno-affinity chromatography is standard for antibody purification.
Purpose of the Study:
- To inform researchers on affinity chromatography techniques for antibody purification.
- To provide guidance on selecting appropriate affinity ligands.
- To cover monoclonal, polyclonal, and recombinant antibody purification.
Main Methods:
- Exploitation of specific binding interactions (ligand-receptor).
- Utilizing affinity chromatography principles.
- Application of immuno-affinity chromatography with Protein A and Protein G.
Main Results:
- Demonstrated the purification of antibodies using affinity chromatography.
- Provided examples for Protein A and Protein G applications.
- Included selection tables for optimal protein ligand choice.
Conclusions:
- Affinity chromatography is a versatile purification method.
- Proteins A and G are key ligands for antibody purification.
- This chapter serves as a practical guide for researchers.
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