Electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry of large biomolecules in complex buffer systems
- Pacific Northwest Lab., Richland, WA (United States)
- Univ.of Texas, Austin, TX (United States)
- Penn State Univ., University Park, PA (United States)
Recent advances in electrospray ionization (ESI) Fourier transform ion cyclotron resonance (FTICR) mass spectrometry allow ultra-high resolution, precision mass measurements of a wide range of biochemically relevant species including proteins, glycoproteins, DNA, and non-covalent complexes. One of the most common difficulties encountered when analyzing biological samples derived from multi-step extraction procedures is obtaining an {open_quotes}electrosprayable{close_quotes} analyte solution which keeps the protein(s) of interest in solution and which does not create numerous background ions which overwhelm the analyte signal or adversely affect the electrospray ionization process. In addition to loss of sensitivity, ionized buffer ions introduce undesirable space charge to the trapped ion cell which limits dynamic range and adversely affects mass measurement precision. Thus, it is highly desirable to minimize the number of background ions in the trapped ion cell. In this poster the authors will present two distinct, and potentially complimentary, techniques which allow high performance mass analysis of biomolecules from complex buffer systems. The first technique, which is particularly well suited for larger analytes, discriminates against low mass matrix ions by the combination of kinetic energy dependent ion accumulation and mass dependent ion loss due to collisionally enhanced magnetron radii. An alternative technique, which is particularly applicable to smaller analytes, is based on electrophoretic separations using on-line capillary zone electrophoresis prior to mass analysis. Preliminary results are presented in which two classes of biomolecules were analyzed from complex buffer systems. These examples include large muscle proteins electrosprayed directly from a 5 M urea buffer and mass analysis from whole human erthrocytes extracted from a buffer which mimics physiological conditions.
- DOE Contract Number:
- AC06-76RL01830
- OSTI ID:
- 61894
- Report Number(s):
- CONF-9405234--
- Country of Publication:
- United States
- Language:
- English
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