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常见缓冲液对表面蚀刻的定量分析及其对无标记生物分析准确性的影响。

Quantification of surface etching by common buffers and implications on the accuracy of label-free biological assays.

机构信息

Department of Biomedical Engineering, Boston University, Boston, MA 02215, USA.

出版信息

Biosens Bioelectron. 2012 Jun-Jul;36(1):222-9. doi: 10.1016/j.bios.2012.04.020. Epub 2012 Apr 21.

Abstract

High throughput analyses in biochemical assays are gaining popularity in the post-genomic era. Multiple label-free detection methods are especially of interest, as they allow quantitative monitoring of biomolecular interactions. It is assumed that the sensor surface is stable to the surrounding medium while the biochemical processes are taking place. Using the Interferometric Reflectance Imaging Sensor (IRIS), we found that buffers commonly used in biochemical reactions can remove silicon dioxide, a material frequently used as the solid support in the microarray industry. Here, we report 53 pm to 731 pm etching of the surface silicon oxide over a 12-h period for several different buffers, including various concentrations of SSC, SSPE, PBS, TRIS, MES, sodium phosphate, and potassium phosphate buffers, and found that PBS and MES buffers are much more benign than the others. We observe a linear dependence of the etch depth over time, and we find the etch rate of silicon dioxide in different buffers that ranges from 2.73±0.76 pm/h in 1M NaCl to 43.54±2.95 pm/h in 6×SSC. The protective effects by chemical modifications of the surface are explored. We demonstrate unaccounted glass etching leading to erroneous results with label-free detection of DNA microarrays, and offer remedies to increase the accuracy of quantitative analysis.

摘要

高通量分析在生化分析中在后基因组时代越来越受欢迎。多种无标记检测方法特别有趣,因为它们允许定量监测生物分子相互作用。人们假设在发生生化过程时,传感器表面对周围介质是稳定的。使用干涉反射成像传感器(IRIS),我们发现生化反应中常用的缓冲液可以去除二氧化硅,二氧化硅是微阵列行业中常用的固体支撑材料。在这里,我们报告了在几种不同的缓冲液中,表面氧化硅在 12 小时内被蚀刻 53 pm 至 731 pm,包括不同浓度的 SSC、SSPE、PBS、TRIS、MES、磷酸钠和磷酸钾缓冲液,并且发现 PBS 和 MES 缓冲液比其他缓冲液更温和。我们观察到随着时间的推移,蚀刻深度呈线性依赖关系,并且我们发现不同缓冲液中二氧化硅的蚀刻速率范围从 1M NaCl 中的 2.73±0.76 pm/h 到 6×SSC 中的 43.54±2.95 pm/h。还探索了表面化学修饰的保护作用。我们证明了无标记的 DNA 微阵列检测中未被注意到的玻璃蚀刻会导致错误的结果,并提供了提高定量分析准确性的补救措施。

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