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Determination of different forms of human interferon‐γ in single natural killer cells by capillary electrophoresis with on‐capillary immunoreaction and laser‐induced fluorescence detection
Author(s) -
Zhang Hua,
Jin Wenrui
Publication year - 2004
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200305803
Subject(s) - capillary electrophoresis , laser induced fluorescence , fluorescein isothiocyanate , fluorescence , chemistry , capillary action , chromatography , detection limit , fluorescein , lysis , biochemistry , materials science , optics , physics , composite material
A novel method for determining different forms of human interferon‐γ (IFN‐γ) in single natural killer cells was developed by capillary electrophoresis (CE) with on‐capillary immunoreaction and laser‐induced fluorescence (LIF) detection. Cells were perforated with digitonin and one single cell was electrokinatically introduced into the front end of a separation capillary. The monoclonal antibody labeled with fluorescein isothiocyanate of IFN‐γ was hydrodynamically injected into the front end of the capillary around the cell introduced. After the cell was lysed by ultrasonication, the front end of the capillary was used as a microreactor to allow different forms of IFN‐γ to process the immunoreaction with their labeled antibody. Finally, the complexes of different forms of IFN‐γ with their labeled antibody were separated and detected by CE with LIF detection with a limit of detection of zeptomoles (10 −21 mol).