期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1104, 期 1-2, 页码 346-351出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2005.11.083
关键词
single-cell analysis; capillary electrophoresis; laser-induced fluorescence detection; immuno-reaction; interferon-gamma
A novel method for single-cell analysis was developed by combining electroporation for intracellular immuno-reaction and capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection. Human interferon-gamma (IFN-gamma) in natural killer (NK) cells was chosen as the test antigen. Two forms of IFN-gamma in single cells could be well separated and detected with a limit of detection of zeptomole. In this assay, the anti-IFN-gamma monoclonal antibody labeled with fluorescein isothiocyanate (Ab*) was introduced into NK cells by electrophoration for intracellular immuno-reaction. After completion of the intracellular immuno-reaction, the NK cells were chemically pre-perforated with digitonin to lyse easily. Then, one NK cell containing the complexes of IFN-gamma isoantigens with Ab* was electrokinetically injected into the capillary. The cell adsorbed on the tip of capillary was lysed by ultrasonication. Finally, the complexes of the different forms of IFN-gamma in the cell were separated and detected by CE-LIF detection. (c) 2005 Elsevier B.V. All rights reserved.
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