A rapid gel electrophoretic chip for serum cholesterol determination.
A rapid gel electrophoretic chip for serum cholesterol determination.
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DOI：
10.1039/c0an00992j
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发表时间：
2011-04
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影响因子：
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通讯作者：
Kenta Kaminikado;Ryuzoh Ikeda;K. Idegami;N. Nagatani;Mun'delanji C. Vestergaard;M. Saito;E. Tamiya
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文献类型：
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作者：
Kenta Kaminikado;Ryuzoh Ikeda;K. Idegami;N. Nagatani;Mun'delanji C. Vestergaard;M. Saito;E. Tamiya
我们提出了一种快速凝胶电泳芯片，由 2.5% (w/v) 丙烯酰胺和 1% (w/v) 琼脂糖凝胶组成，用于使用光刻技术测定血清胆固醇。经过优化后，我们采用常规酶法测定了标准血清的脂蛋白浓度。将血清稀释、染色并加载到芯片上 15 分钟。加载后，使用图像分析仪估计在芯片上分离的低密度脂蛋白胆固醇（LDL-C）和高密度脂蛋白胆固醇（HDL-C）条带的强度。这些条带的强度对应于从标准酶方法获得的浓度。检测到的LDL-C 和HDL-C 浓度分别在146 mg dL(-1) 和53 mg dL(-1) 范围内呈线性。最后，我们使用我们的电泳芯片从九名志愿者获得的真实生物样本进行了胆固醇分析。使用我们的芯片检测到的 LDL-C 和 HDL-C 水平与使用传统酶法获得的结果良好相关，LDL-C 和 HDL-C 的结果分别为 r(2) = 0.98 和 r(2) = 0.86。尽管我们的样本量很小并且仅限于健康志愿者，但我们已经证明这种概念验证凝胶电泳芯片可以同时测定脂蛋白。
We present a rapid gel electrophoretic chip, composed of 2.5% (w/v) acrylamide and 1% (w/v) agarose gel, for serum cholesterol determination using a photo lithography technique. After optimizations, we determined the lipoprotein concentration of standard serum using a conventional enzyme method. The serum was diluted, stained and loaded for 15 min onto the chip. After loading, the intensities of low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDL-C) bands separated at the chip were estimated using an image analyzer. The intensities of these bands corresponded to concentrations obtained from a standard enzyme-based method. The detected LDL-C and HDL-C concentrations were linear up to 146 mg dL(-1) and 53 mg dL(-1) respectively. Finally, we carried out the cholesterol analysis using real biological samples obtained from nine volunteers using our electrophoretic chip. The LDL-C and HDL-C levels detected using our chip correlated well with the results obtained using the conventional enzyme-based method r(2) = 0.98 and r(2) = 0.86 for LDL-C and HDL-C, respectively. Although our sample size is small and confined only to health volunteers, we have demonstrated that this proof-of-concept gel electrophoretic chip can determine lipoproteins, simultaneously.