DEVELOPMENT OF CAPILLARY AND MICROCHIP SEPARATION-DETECTION SYSTEMS TOWARD PROTEOMICS

针对毛细管电泳（CE）和微芯片电泳（MCE）对蛋白质的高效分离和检测，主要研究了以下五个方面。1)建立了一种高性能的微芯片等电聚焦（MCIEF）分析方法。MCIEF在210 s内成功分离了4个模型蛋白，其中蛋白的pI值与其聚焦位置呈良好的线性关系。2)开发了一种固定化蛋白质的高效手性分离技术——毛细管电色谱和微芯片电色谱。以亲和素为手性选择剂，成功地分离了4对对映体。3)在线预富集法提高MCE检测灵敏度。在MCE的胶束电动色谱（MEKC）模式下，利用新设计和制造的T -cross型通道石英芯片，成功地实现了SRW在线预富集。4)利用热透镜显微镜（TLM）提高CE和MCE的检出率。在微芯片MEKC-TLM系统中，以扫描作为在线样品预富集技术，获得了日落黄在4 nM或8 amol绝对注射量下的检出限。5)开发了一种具有质谱（MS）检测的高性能MCE系统。建立了一种具有电喷雾电离界面的MCE-MS系统，成功地分离和检测了部分肽和蛋白质。

Toward the high performance separation and detection of proteins by capillary electrophoresis(CE) and microchip electrophoresis(MCE), following five subjects were mainly investigated.1)Development of a high performance analysis method by microchip isoelectric focusing(MCIEF). Four model proteins were successfully separated within 210 s by MCIEF, where good linear relationship between a pI value of the protein and its focused position.2)Development of a highly efficient chiral separation technique by capillary electrochromatography and microchip electrochromatography using immobilized proteins. Avidin was used as a chiral selector and four pairs of enantiomers were successfully resolved.3)Improvement of detection sensitivity in MCE using on-line sample preconcentration. Under a micellar electrokinetic chromatography (MEKC) mode in MCE, successful on-line sample preconcentration by SRW was achieved by using a newly designed and fabricated T -cross type channel quartz chip.4)Improvement of detectability in CE and MCE by using thermal lens microscopy (TLM). In a microchip MEKC-TLM system with sweeping as an on-line sample preconcentration technique, the limit of detection of 4 nM or 8 amol in absolute amount injected for Sunset Yellow was obtained.5)Development of a high performance MCE system with mass spectrometric (MS) detection. An MCE-MS system with an electrospray ionization interface was developed, where some peptides and proteins were successfully separated and detected.