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. 2022 Jul 13;14(7):478. doi: 10.3390/toxins14070478

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Purification of ObiDi. Step 2: Protein-A affinity chromatography to get rid of free dianthin. The conjugate obtained from cation exchange chromatography (lane “Elution from IEC”) was applied to the column and elution fractions E1 to E7 were collected. All samples were evaluated by (a) SDS-PAGE and (b) Western blotting with a His-tag-HRP antibody under non-reducing conditions. A conjugate of ObiDi with DAR 1 is visible in elution fractions E3 and E4. Free dianthin was successfully removed.