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Single-step purification of cyclotides using affinity chromatography
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy. Khulna University, Pharmacy Discipline .
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Pharmacy, Department of Medicinal Chemistry, Division of Pharmacognosy.
Polish Academy of Science, W. Szafer Institute of Botany.
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2017 (English)In: Biopolymers, ISSN 0006-3525, E-ISSN 1097-0282, Vol. 108, no 3, e23010Article in journal (Refereed) Published
Abstract [en]

Cyclotides are considered promising scaffolds for drug development owing to their inherent host defence activities and highly stable structure, defined by the cyclic cystine knot. These proteins are expressed as complex mixtures in plants. Although several methods have been developed for their isolation and analysis, purification of cyclotides is still a lengthy process. Here, we describe the use of affinity chromatography for the purification of cyclotides using polyclonal IgG antibodies raised in rabbits against cycloviolacin O2 and immobilized on NHS-activated Sepharose columns. Cycloviolacin O2 was used as a model substance to evaluate the chromatographic principle, first as a pure compound and then in combination with other cyclotides, that is, bracelet cyclotide cycloviolacin O19 and Mobius cyclotide kalata B1, and in a plant extract. We demonstrate that single-step purification of cyclotides by affinity chromatography is possible but cross reactivity may occur between homologue cyclotides of the bracelet subfamily.

Place, publisher, year, edition, pages
2017. Vol. 108, no 3, e23010
Keyword [en]
affinity chromatography, cyclotides, cycloviolacin O2, immobilized IgG
National Category
Biochemistry and Molecular Biology Biophysics
Identifiers
URN: urn:nbn:se:uu:diva-334107DOI: 10.1002/bip.23010ISI: 000407992300005OAI: oai:DiVA.org:uu-334107DiVA: diva2:1160014
Funder
Swedish Research Council, 2012-5063
Available from: 2017-11-24 Created: 2017-11-24 Last updated: 2017-11-24Bibliographically approved

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Muhammad, TajShafiullah, MdGöransson, Ulf

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