Retardation Signal for Fluorescent Determination of Total Protein Content via Rapid and Sensitive Chip Moving Reaction Boundary Electrophoretic Titration

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• 作者：Houyu Wang ; Yongting Shi ; Jian Yan ; Jingyu Dong ; Si Li ; Hua Xiao ; Haiyang Xie ; Liu-Yin Fan ; Cheng-Xi Cao
• 刊名：Analytical Chemistry
• 出版年：2014
• 出版时间：March 18, 2014
• 年：2014
• 卷：86
• 期：6
• 页码：2888-2894
• 全文大小：527K
• 年卷期：v.86,no.6(March 18, 2014)
• ISSN：1520-6882

文摘

A novel concept and theory of moving reaction boundary (MRB) retardation signal (R_MRB) was advanced for determination of total protein content via MRB electrophoretic titration (MRBET). The theoretical results revealed that the retardation extent of boundary displacment, viz., the R_MRB value, was as a function of protein content. Thus, the R_MRB value of a sample could be used to determine its total protein content according to the relevant calibration curve. To demonstrate the concept and theoretical results, a novel microdevice was designed for the relevant experiments of MRBET. The microdevice has 30 identical work cells, each of which is composed of five ultrashort single microchannels (5 mm). In the microdevice, fluorescein isothiocyanate (FITC) was used to denote MRB motion and R_MRB value for the first time, the polyacrylamide gel (PAG) containing protein sample was photopolymerized in microchannels, and the MRB was created with acid or alkali and target protein sample. As compared to the classic Kjeldahl method and conventional MRBET performed in glass tube, the developed titration chip has the following merits: good sensitivity (0.3鈥?.4 渭g/mL vs 150鈥?00 渭g/mL of protein concentration, 0.6鈥?.8 ng vs 30鈥?000 渭g of absolute protein content), rapid analysis (20鈥?0 s vs 15鈥?00 min), and portable low-power (15 V vs 200 V).