Abstract
The purification of peptide antibodies (e.g., IgG, IgY, scFv, and Fab) are described in this chapter. Affinity chromatographic purification, a very convenient and effective antibody purification strategy, is used to isolate peptide antibodies based on specific binding, i.e., binding of the antibody to a column on which its specific ligand is immobilized with subsequent elution of the purified antibody. In addition, the application of purification methods based on the use of proteins A, G, and L, each of which bind to specific domains on an antibody/fragment, or the use of specific tags (e.g., histidine and biotin) attached to antibodies or antigens are also described.
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References
Arora S, Ayyar BV, O’Kennedy R (2014) Affinity chromatography for antibody purification. Methods Mol Biol 1129:497–516
McMahon MJ, O’Kennedy R (2000) Polyreactivity as an acquired artefact, rather than a physiologic property, of antibodies: evidence that monoreactive antibodies may gain the ability to bind to multiple antigens after exposure to low pH. J Immunol Methods 241:1–10
Darcy E, Leonard P, Fitzgerald J, Danaher M, O’Kennedy R (2011) Purification of antibodies using affinity chromatography. Methods Mol Biol 681:369–382
Lund LN, Gustavsson PE, Michael R, Lindgren J, Nørskov-Lauritsen L, Lund M, Houen G, Staby A, St Hilaire PM (2012) Novel peptide ligand with high binding capacity for antibody purification. J Chromatogr A 1225:158–167
Sauer-Eriksson AE, Kleywegt GJ, Uhlén M, Jones TA (1995) Crystal structure of the C2 fragment of streptococcal protein G in complex with the Fc domain of human IgG. Structure 3:265–278
Grodzki AC, Berenstein E (2010) Antibody purification: affinity chromatography – protein A and protein G Sepharose. Methods Mol Biol 588:33–41
Nilson BH, Lögdberg L, Kastern W, Björck L, Akerström B (1993) Purification of antibodies using protein L-binding framework structures in the light chain variable domain. J Immunol Methods 164:33–40
Fitzgerald J, Leonard P, Darcy E, O’Kennedy R (2011) Immunoaffinity chromatography. Methods Mol Biol 681:35–59
Ma H, Meng J, Wang J, Hearty S, Dolly JO and O’Kennedy R (2014) Targeted delivery of a SNARE protease to sensory neurons using a single chain antibody (scFv) against the extracellular domain of P2X3 inhibits the release of a pain mediator. Biochem J 462:247–256
Edupuganti SR, Edupuganti OP, O’Kennedy R, Defrancq E, Boullanger S (2013) Use of T-2 toxin-immobilized amine-activated beads as an efficient affinity purification matrix for the isolation of specific IgY. J Chromatogr B Analyt Technol Biomed Life Sci 923–924:98–101
Acknowledgements
This work is supported by Science Foundation Ireland under CSET Grant No. 05/CE3/B754 and 10/CE/B1821.
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Ma, H., O’Kennedy, R. (2015). The Purification of Natural and Recombinant Peptide Antibodies by Affinity Chromatographic Strategies. In: Houen, G. (eds) Peptide Antibodies. Methods in Molecular Biology, vol 1348. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2999-3_15
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DOI: https://doi.org/10.1007/978-1-4939-2999-3_15
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-2998-6
Online ISBN: 978-1-4939-2999-3
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