Abstract
Multimodal (MM) chromatography can be described as a chromatographic method that uses more than one mode of interaction between the target molecule and the ligand to achieve a particular separation. Owing to its advantages over traditional chromatography, such as higher selectivity and capacity, its application for the purification of biomolecules with therapeutic interest has been widely studied. The potential of MM chromatography for the purification of plasmid DNA has been demonstrated. In this chapter, a downstream process for the purification of supercoiled plasmid DNA using MM chromatography with two different ligands—Capto™ adhere and PPA HyperCell™—is described. In both the cases, the purification process yields a high purity and highly homogeneous sc plasmid product.
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References
Sun B, Yu X, Yin Y et al (2013) Large-scale purification of pharmaceutical-grade plasmid DNA using tangential flow filtration and multi-step chromatography. J Biosci Bioeng 116:281–286
Halan V, Maity S, Bhambure R et al (2019) Multimodal chromatography for purification of biotherapeutics – a review. Curr Protein Pept Sci 20:4–13
Yang Y, Geng X (2011) Mixed-mode chromatography and its applications to biopolymers. J Chromatogr A 1218:8813–8825
Kallberg K, Johansson H-O, Bulow L (2012) Multimodal chromatography: an efficient tool in downstream processing of proteins. Biotechnol J 7:1485–1495
Pinto IF, Aires-Barros MR, Azevedo AM (2015) Multimodal chromatography: debottlenecking the downstream processing of monoclonal antibodies. Pharm Bioprocess 3:263–279
Zhao G, Dong X-Y, Sun Y (2009) Ligands for mixed-mode protein chromatography: principles, characteristics and design. J Biotechnol 144:3–11
Silva-Santos AR, Alves CPA, Prazeres DMF et al (2016) Separation of plasmid DNA topoisomers by multimodal chromatography. Anal Biochem 503:68–70
Sousa A, Pereira P, Sousa F et al (2014) Binding mechanisms for histamine and agmatine ligands in plasmid deoxyribonucleic acid purifications. J Chromatogr A 1366:110–119
Silva-Santos AR, Alves CP, Prazeres DMF et al (2017) A process for supercoiled plasmid DNA purification based on multimodal chromatography. Sep Purif Technol 182:94–100
Matos T, Queiroz JA, Bülow L (2014) Plasmid DNA purification using a multimodal chromatography resin. J Mol Recognit 27:184–189
Černigoj U, Vidic U, Barut M et al (2013) A multimodal histamine ligand for chromatographic purification of plasmid DNA. J Chromatogr A 1281:87–93
Prazeres DMF (2011) Plasmid biopharmaceuticals: basics, applications, and manufacturing. John Wiley & Sons, Hoboken, NJ. , 615p. https://doi.org/10.1002/9780470939918.ch12
Brenac BV, Schapman A, Santambien P et al (2008) Fast purification process optimization using mixed-mode chromatography sorbents in pre-packed mini-columns. J Chromatogr A 1177:226–233
Ghanem A, Healey R, Adly FG (2013) Current trends in separation of plasmid DNA vaccines: a review. Anal Chim Acta 760:1–15
Gonçalves GAL, Prather KLJ, Monteiro GA et al (2014) Plasmid DNA production with Escherichia coli GALG20, a pgi-gene knockout strain: fermentation strategies and impact on downstream processing. J Biotechnol 186:119–127
Acknowledgments
Funding received by iBB—Institute for Bioengineering and Biosciences from FCT–Portuguese Foundation for Science and Technology, Portugal (grant UID/BIO/04565/2019) and from Programa Operacional Regional de Lisboa 2020, Portugal (Project N. 007317) is acknowledged. A. Rita Silva-Santos acknowledges FCT for the PhD grant PD/BD/135138/2017 (BIOTECnico program). This work was supported by National Funds by FCT—Fundação para a Ciência e Tecnologia—through the research contract IF/00048/2014/CP1214/CT0010.
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Silva-Santos, A.R., Rosa, S.S., Prazeres, D.M.F., Azevedo, A.M. (2021). Purification of Plasmid DNA by Multimodal Chromatography. In: Sousa, Â. (eds) DNA Vaccines. Methods in Molecular Biology, vol 2197. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0872-2_10
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DOI: https://doi.org/10.1007/978-1-0716-0872-2_10
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