Application of capillary electrophoresis in analysis of intact mammalian cells

Transcript

1 Vol. 30 No. 2 February 2012 Chinese Journal of Chromatography 116 ~ 122 DOI /SP. J * O658 A Application of capillary electrophoresis in analysis of intact mammalian cells ZHANG Lu QU Feng * LOU Beilei School of Life Science Beijing Institute of Technology Beijing China Abstract Cell is the basic structural and functional unit of human body. The research of cells structure function and behavior is very important. Capillary electrophoresis CE is a pow erful tool for the separation and analysis the application of w hich in cell analysis has progressed significantly. In this paper the developments of CE applied in the intact mammalian cell analysis are review ed w hich consist of cell population and single cell analysis. The erythrocyte boar sperm HeLa cells SH-SY5Y cells Caco -2 cells K562 cells and rat cerebellar granule cells are invo lved in this review. The m etho ds and co nditio ns fo r the intact m am m alian cell analysis are summarized. In addition the problems caused by the breakage aggregation sedimentation adsorption and electrophoretic heterogeneity of the cell in the intact mammalian cell analysis by CE are discussed and the corresponding solutions are introduced. Also the future research trends are presented. Fo rty nine papers in all are review ed. Key words capillary electrophoresis CE mammalian cells analysis of intact cells CE CE CE *. Tel qufengqu@ bit. edu. cn. Nos No. 2012CB

2 2 117 CE Chen 22 CZE mm min min min 14 min 19 ph CZE Lu 23 CE CE CE CE 10 ~ 200 μm 7 ~ 200 μm CE CE CE 21 7 CE CE CE-UV-MVI CE CZE Torimura 27 25

3 CZE - CE-LIF 33 CZE ph HeLa B / green /red fluorescent signal G /R HeLa CE-LIF - UV-Vis 214 nm CE Gao 32 Armstrong 28 SH-SY5Y SYBER- 14 PI charge coupled device CCD ~ 596 nm B CE methylthiazolyl- bromide MTT diphenyl-tetrazolium CZE DNA Ru 29 CE HeLa DNA 3 AO 34 Caco -2 CZE DAD UV-Vis 30 R 590 /214 R 590 /214 HeLa / Guo methyl-4-phenyl pyridinium ion MPP MPP +

4 % Tsuda K562 HeLa CE 40 Xiao 44 CE-LIF CE K562 P- P-glycoprotein PGP 2. 1 Kitagaw a μm 89 PGP CZE 35 CZE CE Tsuda 42 CE CZE HeLa 25 μm CZE 100 CE 20 mau 10 mau Tsuda HeLa nm 590 nm G PGP /mol 10 ~ 20 mau G

5 CE MTT CE 1 Cell Fixing agent Erythrocyte 3% formalin or 1% glutaraldehyde Table 1 1 CE Methods and conditions for intact cell analysis by CE Coating material of capillary - Additive in electrophoresis buffer 0. 1% % hydroxypropyl methyl cellulose Detector Publication date Reference UV Erythrocyte - - 6% sucrose UV Erythrocyte UV microscopy Erythrocyte - - 6% sucrose UV Erythrocyte g / L hydroxypropyl methyl cellulose Erythrocyte % hydroxypropyl methyl cellulose UV UV Boar sperm % % PEO LIF HeLa UV-Vis HeLa UV Rat cerebellar granule cells UV-Vis SH-SY5Y UV-Vis HeLa 4% formaldehyde - - DAD Caco -2 4% paraformaldehyde - - DAD - 34 Erythrocyte - - serum microscopy Erythrocyte - bovine serum albumin 1% bovine serum albumin or % agar or 10% glucose UV-Vis microscopy Erythrocyte - human serum - microscopy K562 - polyacrylamide - LIF HeLa 4% formaldehyde - - UV-Vis CE CE h

6 K562 /S RSD 24% ~ 87% CE CE 49 Guo 31 RSD 10. 8% RSD % % ~ 0. 2% HPMC CE HPMC CE CE 26 BSA electrophoretic heterogeneity K562 /A PGP RSD 20% ~ 122% 1 Jorgenson J W Lukacs K D. Science Gotti R. J Pharmaceut Biomed Guihen E O Connor W T. Electrophoresis Guzman N A Phillips T M. Electrophoresis Bakry R Huck C W Najam-ul-Haq M et al. J Sep Sci P Dawod M Chung D S. J Sep Sci Herrero M Iba ez E Cifuentes A. Electrophoresis Qu F. Chinese Journal of Chromatography Subirats X Blaas D Kenndler E. Electrophoresis Kremser L Bilek G Blaas D et al. J Sep Sci

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