Abstract
The availability of purified antibodies is prerequisite for many applications and the appropriate choice(s) of antibody-purification steps is crucial. Numerous methods have been developed for the purification of antibodies; however, affinity chromatography-based methods are the most extensively utilized. These methods are based on highly specific and reversible biological interactions between two molecules (e.g., between receptor and ligand or antibody and antigen). Affinity chromatography offers very high selectivity, involving minimal steps, providing simplicity of approach and rapidity. Implementing an effective protocol often requires meticulous planning and testing in order to achieve high purity and yields of desired antibody types/subtypes. This chapter describes the basic techniques for purification of monoclonal, polyclonal, and recombinant antibodies employing affinity chromatography.
Key words
- Antibody purification
- Affinity chromatography
- Monoclonal antibodies
- Polyclonal antibodies
- Recombinant antibodies
- Protein A
- Protein G
- Protein L
- Thiophilic chromatography
- Immobilized metal affinity chromatography
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Acknowledgments
We gratefully acknowledge the support of Science Foundation Ireland (CSET Grant numbers 05/CE3/B754 and 10/CE/B1821); the Irish Research Council for Science, Engineering and Technology (IRCSET) Embark Scholarship (to Sushrut Arora); the Biomedical Diagnostics Institute (BDI); and the National Centre for Sensor Research (NCSR).
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Arora, S., Ayyar, B.V., O’Kennedy, R. (2014). Affinity Chromatography for Antibody Purification. In: Labrou, N. (eds) Protein Downstream Processing. Methods in Molecular Biology, vol 1129. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-977-2_35
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DOI: https://doi.org/10.1007/978-1-62703-977-2_35
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