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Acquisition of enzymatic progress curves in real time by quenching-free ion exchange chromatography

Agustoni, Elia and Teixeira, Raphael Dias and Huber, Markus and Flister, Susanne and Hiller, Sebastian and Schirmer, Tilman. (2022) Acquisition of enzymatic progress curves in real time by quenching-free ion exchange chromatography. Analytical biochemistry, 639. p. 114523.

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Official URL: https://edoc.unibas.ch/85935/

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Abstract

We describe a quenching-free, 'online' ion exchange chromatography (oIEC) method for the quantitative analysis of enzymatic reactions in real-time. We show that separate quenching of the ongoing reaction performed conventionally is not required, since enzymatic reactions are interrupted upon immobilization of the reaction compounds by binding to the stationary phase of the ion exchange column. The reaction mix samples are directly injected into the column, thereby improving data consistency and allowing automation of the process. The method allows reliable and efficient acquisition of enzymatic progress curves by automatic loading of aliquots of an ongoing reaction at predefined timepoints. We demonstrate the applicability of this method for a variety of enzymatic reactions. SUBJECT: Enzymatic assays and analysis.
Faculties and Departments:05 Faculty of Science > Departement Biozentrum > Structural Biology & Biophysics > Structural Biology (Hiller)
05 Faculty of Science > Departement Biozentrum > Former Organization Units Biozentrum > Structural Biology (Schirmer)
UniBasel Contributors:Schirmer, Tilman and Agustoni, Elia and Dias Teixeira, Raphael and Huber, Markus and Flister, Susanne and Hiller, Sebastian
Item Type:Article, refereed
Article Subtype:Research Article
Publisher:Elsevier
ISSN:1096-0309
Note:Publication type according to Uni Basel Research Database: Journal article
Identification Number:
Last Modified:27 Jan 2022 13:47
Deposited On:27 Jan 2022 13:47

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