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使用微芯片电泳和质谱分析凝血酶-抗凝血酶复合物的形成。

Analysis of thrombin-antithrombin complex formation using microchip electrophoresis and mass spectrometry.

机构信息

Department of Chemistry and Biochemistry, Brigham Young University, Provo, UT, USA.

出版信息

Electrophoresis. 2019 Nov;40(21):2853-2859. doi: 10.1002/elps.201900235. Epub 2019 Aug 13.

Abstract

Preterm birth (PTB) related health problems take over one million lives each year, and currently, no clinical analysis is available to determine if a fetus is at risk for PTB. Here, we describe the preparation of a key PTB risk biomarker, thrombin-antithrombin (TAT), and characterize it using dot blots, MS, and microchip electrophoresis (µCE). The pH for fluorescently labeling TAT was also optimized using spectrofluorometry and spectrophotometry. The LOD of TAT was measured in µCE. Lastly, TAT was combined with six other PTB risk biomarkers and separated in µCE. The ability to make and characterize TAT is an important step toward the development of an integrated microfluidic diagnostic for PTB risk.

摘要

早产(PTB)相关的健康问题每年导致超过 100 万人死亡,目前尚无临床分析可确定胎儿是否有早产风险。在这里,我们描述了一种关键的 PTB 风险生物标志物——凝血酶-抗凝血酶(TAT)的制备,并使用点印迹、MS 和微芯片电泳(µCE)对其进行了表征。还使用荧光分光光度法和分光光度法优化了 TAT 的荧光标记 pH 值。在 µCE 中测量了 TAT 的检测限。最后,将 TAT 与其他六个 PTB 风险生物标志物结合并在 µCE 中分离。制备和表征 TAT 的能力是开发用于 PTB 风险的集成微流控诊断的重要步骤。

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Microchip electrophoresis separation of a panel of preterm birth biomarkers.微流控芯片电泳分离一组早产生物标志物。
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本文引用的文献

1
Blood compatible materials: state of the art.血液相容性材料:当前技术水平
J Mater Chem B. 2014 Sep 21;2(35):5718-5738. doi: 10.1039/c4tb00881b. Epub 2014 Aug 1.
4
Microchip electrophoresis separation of a panel of preterm birth biomarkers.微流控芯片电泳分离一组早产生物标志物。
Electrophoresis. 2018 Sep;39(18):2300-2307. doi: 10.1002/elps.201800078. Epub 2018 Jun 1.
9
Universal sample preparation method for proteome analysis.蛋白质组分析的通用样本制备方法。
Nat Methods. 2009 May;6(5):359-62. doi: 10.1038/nmeth.1322. Epub 2009 Apr 19.

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