کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1203003 1493574 2014 7 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Capillary electrophoresis-based immobilized enzyme reactor using particle-packing technique
ترجمه فارسی عنوان
با استفاده از روش بسته بندی ذرات، راکتور آنزیم مجهز به الکتروفورز موثر است
کلمات کلیدی
میکروسکوپ آنزیم، بسته بندی ذرات، الکتروفورز مویرگی، تریپسین هضم
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
چکیده انگلیسی


• We applied packing technique to fabricate CE-based IMER using single particle as the frits.
• It takes less than 5 min to fabricate a 5-mm long IMER using the method.
• The microreactor exhibits excellent reproducibility and stability for analysis of enzyme reaction.
• The method expands the application of capillary electrophoresis for on-line enzyme assay.

A novel method using particle-packing technique to fabricate capillary electrophoresis (CE)-based immobilized enzyme reactor (IMER) was accomplished by utilizing perfusive silica single particles as the frits and large-pore beads as the enzyme supports. The fabrication procedure is rapid and simple; the length and enzyme loading amount of the CE-IMERs could be easily adjusted. Performance and feasibility of the CE-IMERs were investigated using on-line trypsin digestion as the model enzyme reaction. High reproducible on-line enzyme assay was demonstrated with RSD less than 4.1% and 3.8% for peak area and migration time of the substrate and product over 100 consecutive runs, respectively. The enzyme can still maintain the activity for at least 10 days, indicating remarkably stability of the CE-IMERs. The CE-IMERs were successfully applied for accurate analysis of trypsin inhibition as well as on-line digestion of standard proteins (myoglobin and BSA). The present method provides a new interesting alternative to open-tubular and monolithic CE-IMERs, thus expands the application of the CE technique for on-line enzyme assay and analysis and characterization of peptides and proteins.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Journal of Chromatography A - Volume 1352, 25 July 2014, Pages 80–86
نویسندگان
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