Direct analysis of cellular proteins by capillary electrophoresis FTICR MS
- Pacific Northwest Lab., Richland, WA (United States); and others
Direct chemical analysis of living cells has received considerable attention in recent years; the single cell approach provides a major step towards answering important questions in the field of cellular biochemistry. In this work, the authors present preliminary results which demonstrate the feasibility of using the CE-ESI-FTICR combination as a high performance detection scheme for the analysis of cellular proteins acquired directly from small populations of intact living cells. The human erythrocyte (red blood cell) was chosen as a model system owing to its availability, relatively homogeneous composition, and thorough documentation of contents by previous researchers. The contents of the erythrocyte are unusually homogeneous; nearly the entire volume of the cell is filled with hemoglobin, approximately 450 amol per cell, a challenging but attainable level for mass spectrometric detection with current instrumentation. In this work, the authors demonstrate the on-line acquisition of high resolution mass spectra (average resolution {ge} 45,000 FWHM) of the {alpha} and {Beta} hemoglobin chains acquired from the injection of as few as 10 human erythrocytes (this corresponds to {approx} 4.5 fmol of hemoglobin). Additionally, when used in conjunction with quadrupolar axialization and sustained off-resonance irradiation, it is possible to directly obtain partial sequence information of selected cellular components obviating the need for additional isolation/purification steps. Given the extremely small volume of the human erythrocyte (typically {approx} 87 fL/cell), the authors are optimistic that the techniques implemented here will be adaptable to the study of many larger mammalian cell systems.
- OSTI ID:
- 210626
- Report Number(s):
- CONF-9505261--
- Country of Publication:
- United States
- Language:
- English
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