Microchip electrophoresis of N-glycans on serpentine separation channels with asymmetrically tapered turns.
Microchip electrophoresis of N-glycans on serpentine separation channels with asymmetrically tapered turns.
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DOI：
10.1002/elps.201000461
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发表时间：
2011-01
期刊：
影响因子：
2.9
通讯作者：
Jacobson, Stephen C.
中科院分区：
文献类型：
--
作者：
Zhuang, Zexi;Mitra, Indranil;Hussein, Ahmed;Novotny, Milos V.;Mechref, Yehia;Jacobson, Stephen C.
我们设计和制造了具有蛇形分离通道和不对称锥形转角的微流控器件，从而实现了高效分离，并最大限度地减少了与“赛道”效应相关的频带展宽。我们通过测量分离效率随分离长度、电场强度、匝数锥度比和匝数的变化来评估这些器件的性能。以8-氨基比林-1，3，6-三磺酸标记的核糖核酸酶B(RNaseB)衍生的N-葡聚糖在蛇形通道上进行了分离，分离长度分别为11、18、22和36 cm，电场强度为750~1750V/cm。36厘米通道上的分离产生的板数高达940,000个，分析时间不到3.1分钟，而22厘米通道上的分离具有较短的分析时间(不到1.25分钟)，仍然具有相当的效率(高达600,000个板)。当锥度比为2和3时，转弯诱导的色散最小，而沿分离长度有两个或四个180°转弯对整体效率没有影响。所开发的装置被用于分析来自卵巢癌患者和健康人血清的天然和所需的N-糖链。对于这些生物样品，获得了与从核糖核酸酶B中分离模型多糖相似的分离效率。
We designed and fabricated microfluidic devices with serpentine separation channels and asymmetrically tapered turns, thus allowing high efficiency separations and minimizing band broadening associated with the “racetrack” effect. We evaluated the performance of these devices by measuring the variation in separation efficiency with separation length, electric field strength, taper ratio of the turns, and number of turns. N-Glycans derived from ribonuclease B (RNase B) and labeled with 8-aminopyrene-1,3,6-trisulfonic acid were electrophoretically separated on serpentine channels with separation lengths of 11, 18, 22, and 36 cm at electric field strengths from 750 to 1750 V/cm. Separations on the 36-cm channel produced plate numbers up to 940,000 with an analysis time under 3.1 min, whereas separations on the 22-cm channel had a shorter analysis time (less than 1.25 min), still with respectable efficiencies (up to 600,000 plates). Turn-induced dispersion was minimized with taper ratios 2 and 3, whereas having two or four 180° turns along the separation length did not impact the overall efficiency. The developed device was used to analyze native and desialylated N-glycans derived from the blood serum of an ovarian cancer patient and a disease-free individual. Separation efficiencies similar to that achieved with the model glycans from RNase B were attained for these biological samples.
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