کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1199591 1493554 2014 8 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Thermodynamic study of the interaction between linear plasmid deoxyribonucleic acid and an anion exchange support under linear and overloaded conditions
ترجمه فارسی عنوان
بررسی ترمودینامیکی تعامل بین اسید پراکسید ریبونوکلئیک اسید و حمایت تبادل آنی تحت شرایط خطی و پر بار
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• Linear plasmid DNA adsorption process enthalpically driven for studied anion exchanger.
• Profound role of desolvation process in the adsorption mechanism.
• Plasmid DNA upright position preferred when binding under maximum loading conditions.
• Plasmid DNA reorientation on the adsorbent surface at linear and overloaded conditions.

Anion-exchange chromatography has been successfully used in plasmid DNA (pDNA) purification. However, pDNA adsorption mechanism using this method is still not completely understood, and the prediction of the separation behavior is generally unreliable. Flow microcalorimetry (FMC) has proven its ability to provide an improved understanding of the driving forces and mechanisms involved in the adsorption process of biomolecules onto several chromatographic systems. Thus, using FMC, this study aims to understand the adsorption mechanism of linear pDNA (pVAX1-LacZ) onto the anion-exchange support Fast Flow (FF) Q-Sepharose. Static binding capacity studies have shown that the mechanism of pDNA adsorption onto Q-Sepharose follows a Langmuir isotherm. FMC experiments resulted in thermograms that comprised endothermic and exothermic heats. Endothermic heat major contributor was suggested to be the desolvation process. Exothermic heats were related to the interaction between pDNA and Q-Sepharose primary and secondary adsorption. Furthermore, FMC revealed that the overall adsorption process is exothermic, as expected for an anion-exchange interaction. Nevertheless, there are evidences of the presence of nonspecific effects, such as reorientation and electrostatic repulsive forces.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1372, 12 December 2014, Pages 166–173
نویسندگان
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