Novel sheathless CE-MS interface as an original and powerful infusion platform for nanoESI study: from intact proteins to high molecular mass noncovalent complexes

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• 作者：Rabah Gahoual (1)
  Jean-Marc Busnel (2)
  Philippe Wolff (3)
  Yannis Nicolas Fran?ois (1)
  Emmanuelle Leize-Wagner (1)
  
• 关键词：Sheathless capillary electrophoresis ; Mass spectrometry ; Noncovalent complexes
• 刊名：Analytical and Bioanalytical Chemistry
• 出版年：2014
• 出版时间：February 2014
• 年：2014
• 卷：406
• 期：4
• 页码：1029-1038
• 全文大小：277 KB
• 作者单位：Rabah Gahoual (1)
  Jean-Marc Busnel (2)
  Philippe Wolff (3)
  Yannis Nicolas Fran?ois (1)
  Emmanuelle Leize-Wagner (1)
  
  1. Laboratoire de Spectrométrie de Masse des Interactions et des Systèmes (LSMIS), CNRS-UMR 7140, Université de Strasbourg, 6700, Strasbourg, France
  2. Beckman Coulter Inc., Brea, CA, 92821, USA
  3. Architecture et Réactivité de l’ARN, Institut de Biologie Moléculaire et Cellulaire, Université de Strasbourg, Strasbourg, France
  
• ISSN：1618-2650

文摘

Development of nano-electrospray (nanoESI) sources allowed to increase significantly the sensitivity which is often lacking when studying biological noncovalent assemblies. However, the flow rate used to infuse the sample into the mass spectrometer cannot be precisely controlled with nanoESI and the robustness of the system could represent an issue. In this study, we have used a sheathless capillary electrophoresis–mass spectrometry (CESI) prototype as a nanoESI infusion device. The hydrodynamic mobilization of the capillary content was characterized and the ability of the system to generate a stable electrospray under controlled flow rate conditions ranging from 4 up to 900?nL/min was demonstrated. The effect of the infusing flow rate on the detection of an intact model protein analyzed under native conditions was investigated. Results demonstrated a significant increase in sensitivity of 46-fold and a signal-to-noise ratio improvement of nearly 5-fold when using an infusing flow rate from 456.9 down to 13.7?nL/min. The CESI prototype was further used to detect successfully the β ring homodimer in its native conformation. Obtained results were compared with those achieved with conventional ESI. Intensity signals were increased by a factor of 5, while sample consumption decreased 80 times. β ring complexed with the P14 peptide was also studied. Finally, the CESI interface was used to observe the quaternary structure of native hemocyanins from Carcinus maenas crabs; this high molecular complex coexisting under various degrees of complexation and resulting in masses ranging from 445?kDa to 1.34?MDa.