title: A simple heated-capillary modification improves the analysis of non-covalent complexes by Z-spray electrospray ionization.
abstract: BACKGROUND
  The observation of intact non-covalent complexes by electrospray ionization mass spectrometry (ESI-MS) hinges on the ability to minimize in-source activation processes that take place during analyte desolvation. We explored the merits of replacing the sampling cone of a standard Z-spray source with a heated capillary that makes the desolvation process slower and more gradual. We employed well-characterized protein-RNA, RNA-RNA, and DNA-DNA assemblies to compare the alternative configurations.
  
  
  METHODS
  Mass analysis evaluated the integrity of the complexes, whereas traveling wave ion mobility experiments assessed the stability of biomolecular structure. Analyses were performed back-to-back on the same samples on a Synapt G2 HDMS equipped with either the standard sampling cone or the heated-capillary apparatus. In each configuration, the source/capillary temperature was varied in controlled fashion, while keeping all other desolvation parameters constant to monitor the in-source dissociation of selected DNA duplexes. Ion mobility data were obtained from the same precursor by using the alternative configurations under the same settings.
  
  
  RESULTS
  Monitoring the percentage of associated complex demonstrated that the heated capillary provided softer desolvation that was more conducive to the detection of intact non-covalent interactions. This configuration failed to produce complete dissociation of 14 bp and 24 bp duplexes, even when the source/capillary temperature was increased well above their solution melting points. Analyzed by IMS-MS, a selected construct displayed just one conformation with the heated capillary, but two with the standard sampling cone.
  
  
  CONCLUSIONS
  The heated capillary minimizes in-source activation processes that can lead to unintended dissociation of complexes and perturbation of biomolecular structure, which rely on the integrity of non-covalent interactions. This effect can be attributed to the attenuation of the supersonic expansion typical of the Z-spray geometry and the greater ability to control the energy imparted to the system. This hardware modification will be expected to benefit the analysis of biomolecular structure performed on this particular instrumental platform.
authors: The RNA Institute, University at Albany (SUNY), Life Sciences Research Building, Room 1109, 1200 Washington Ave, Albany, NY, 12222, USA.
authors: Jennifer L
authors: JL
authors: Lippens
authors: The RNA Institute, University at Albany (SUNY), Life Sciences Research Building, Room 1109, 1200 Washington Ave, Albany, NY, 12222, USA.
authors: J B
authors: JB
authors: Mangrum
authors: The RNA Institute, University at Albany (SUNY), Life Sciences Research Building, Room 1109, 1200 Washington Ave, Albany, NY, 12222, USA.
authors: William
authors: W
authors: McIntyre
authors: The RNA Institute, University at Albany (SUNY), Life Sciences Research Building, Room 1109, 1200 Washington Ave, Albany, NY, 12222, USA.
authors: Bill
authors: B
authors: Redick
authors: The RNA Institute, University at Albany (SUNY), Life Sciences Research Building, Room 1109, 1200 Washington Ave, Albany, NY, 12222, USA.
authors: D
authors: D
authors: Fabris
mesh_terms: D004867:Equipment Design; D006358:Hot Temperature; D009696:Nucleic Acids / Q000032:analysis / Q000737:chemistry; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation*
chemical_list: D009696:Nucleic Acids