کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1175577 961808 2006 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Integration of an on-line protein digestion microreactor to a nanoelectrospray emitter for peptide mapping
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آنالیزی یا شیمی تجزیه
پیش نمایش صفحه اول مقاله
Integration of an on-line protein digestion microreactor to a nanoelectrospray emitter for peptide mapping
چکیده انگلیسی

A method for integrating nanoelectrospray mass spectrometry with a microreactor for on-line digestion and fast peptide mass mapping from dilute protein samples is presented. Fused silica capillaries (i.d. 50 μm, o.d. 360 μm) are employed as the digestion microreactor and the nanoelectrospray emitter by immobilizing trypsin onto the surface of the inner wall of the fused silica capillary tubing. The procedure is demonstrated using solutions of 1 pmol/μl angiotensin II, cytochrome c, hemoglobin, and β-casein. Because the inner walls of the capillaries are modified by covalent chemical bonds, the adsorption of peptides and proteins to the inner walls of the capillaries is suppressed. This procedure was performed with solutions as dilute as 1 fmol/μl (1 nM) cytochrome c. This method shows generation of tryptic peptides with sequence coverage up to 90% within minutes; trypsin autolysis products are not detected. In addition, the immobilized enzyme can be cleaned easily, enabling the microreactor to be reused for nanoelectrospray.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Analytical Biochemistry - Volume 359, Issue 2, 15 December 2006, Pages 167–175
نویسندگان
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