دورية أكاديمية
Resin structure impacts two-component protein adsorption and separation in anion exchange chromatography.
| العنوان: | Resin structure impacts two-component protein adsorption and separation in anion exchange chromatography. |
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| المؤلفون: | Beck J; Institute of Bioprocess Science and Engineering, Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria., Hochdaninger G; Institute of Bioprocess Science and Engineering, Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria., Carta G; Department of Chemical Engineering, University of Virginia, Charlottesville, VA, USA., Hahn R; Institute of Bioprocess Science and Engineering, Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, Austria. Electronic address: rainer.hahn@boku.ac.at. |
| المصدر: | Journal of chromatography. A [J Chromatogr A] 2023 Aug 30; Vol. 1705, pp. 464208. Date of Electronic Publication: 2023 Jul 12. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 9318488 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1873-3778 (Electronic) Linking ISSN: 00219673 NLM ISO Abbreviation: J Chromatogr A Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Amsterdam ; New York : Elsevier, 1993- |
| مواضيع طبية MeSH: | Conalbumin* , Anion Exchange Resins*/chemistry, Adsorption ; Green Fluorescent Proteins ; Polymers/chemistry ; Chromatography, Ion Exchange ; Kinetics ; Anions |
| مستخلص: | The influence of the resin structure, on the competitive binding and separation of a two-component protein mixture with anion exchange resins is evaluated using conalbumin and green fluorescent protein as a model system. Two macroporous resins, one with large open pores and one with smaller pores, are compared to a resin with grafted polymers. Investigations include measurements of single and two-component isotherms, batch uptake kinetics and two-component column breakthrough. On both macroporous resins, the weaker binding protein, conalbumin, is displaced by the stronger binding green fluorescent protein. For the large pore resin, this results in a pronounced overshoot and efficient separation by frontal chromatography. The polymer-grafted resin exhibits superior capacity and kinetics for one-component adsorption, but is unable to achieve separation due to strongly hindered counter-diffusion. Intermediate separation efficiency is obtained with the smaller pore resin. Confocal laser scanning microscopy provides a mechanistic explanation of the underlying intra-particle diffusional phenomena revealing whether unhindered counter-diffusion of the displaced protein can occur or not. This study demonstrates that the resin's intra-particle structure and its effects on diffusional transport are crucial for an efficient separation process. The novelty of this work lies in its comprehensive nature which includes examples of the three most commonly used resin structures: a small pore agarose matrix, a large-pore polymeric matrix, and a polymer grafted resin. Comparison of the protein adsorption properties of these materials provides valuable clues about advantages and disadvantages of each for anion exchange chromatography applications. Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. (Copyright © 2023 The Author(s). Published by Elsevier B.V. All rights reserved.) |
| فهرسة مساهمة: | Keywords: Batch adsorption; CLSM; Frontal chromatography; Mass transfer |
| المشرفين على المادة: | 147336-22-9 (Green Fluorescent Proteins) 1391-06-6 (Conalbumin) 0 (Anion Exchange Resins) 0 (Polymers) 0 (Anions) |
| تواريخ الأحداث: | Date Created: 20230715 Date Completed: 20230809 Latest Revision: 20230809 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1016/j.chroma.2023.464208 |
| PMID: | 37453173 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1873-3778 |
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| DOI: | 10.1016/j.chroma.2023.464208 |