Capillary Sieving Electrophoresis/Micellar Electrokinetic Capillary Chromatography for Two-Dimensional Protein Fingerprinting of Single Mammalian Cells

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• 作者：Shen Hu ; David A. Michels ; Md. Abul Fazal ; Chootima Ratisoontorn ; Michael L. Cunningham ; and Norman J. Dovichi
• 刊名：Analytical Chemistry
• 出版年：2004
• 出版时间：July 15, 2004
• 年：2004
• 卷：76
• 期：14
• 页码：4044 - 4049
• 全文大小：253K
• 年卷期：v.76,no.14(July 15, 2004)
• ISSN：1520-6882

文摘

We have developed a two-dimensional capillary electrophoresis method for the study of protein expression insingle mammalian cells. The first-dimension capillarycontains an SDS-pullulan buffer system to performcapillary sieving electrophoresis, which separates proteinsbased on molecular weight. The second-dimension capillary contains an SDS buffer for micellar electrokineticcapillary chromatography. After a 6-min-long preliminaryseparation, fractions from the first capillary are successively transferred to a second capillary, where theyundergo further separation by MECC. Over 100 transfersand second-dimension separations are performed over an~3.5-h-long period. We demonstrate this technology bygenerating protein fingerprints from single native MC3T3-E1 osteoprogenitor cells and MC3T3-E1 cells transfectedwith the human transcription regulator TWIST. We alsopresent single-cell protein fingerprints from MCF-7 breastcancer cells before and following treatment to induceapoptosis.