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Following hybridization on sensor/array platforms by using SPR, elipsometer and MALDI-MS
Nucleosides, Nucleotides & Nucleic Acids ( IF 1.3 ) Pub Date : 2020-05-13 , DOI: 10.1080/15257770.2020.1750635
Ömür Çelikbıçak 1 , Kadriye Özlem Hamaloğlu 2 , Bekir Salih 1 , Erhan Pişkin 3
Affiliation  

Abstract The aim of this study is to develop a methodology in which Surface Plasmon Resonance (SPR), Ellipsometer (EM) and Matrix-Assisted Laser Desorption/Ionization-Mass Spectrometry (MALDI-MS) will be used together for detection of single-strand oligodeoxynucleotides (ssODNs) targets. A selected target-ssODNs, and its complementary, the probe-ssODNs carrying a -SH end group, a spacer arm (HS-(CH2)6–(T)15, and a non-complementary ssODNs were used. Silicone based stamps with 16 regions were prepared and used for micro-contact printing (µCP) of the probe-ssODNs on the gold coated surfaces homogeneously. A modulator-spacer molecule (6-mercapto-1-hexanol) was co-immobilized to control surface probe density, to orientate the probe-ssODNs, and to eliminate the nonspecific interactions. SPR was used successfully to follow the hybridization of the target-ssODNs with the immobilized probe-ssODNs on the platform surfaces. Complete hybridizations were achieved in 100 min. It was obtained that there was a linear relationship between relative change in delta and target concentration below 1 µm. Using imaging version of ellipsometer (IEM) allowed imaging of the surfaces and supported extra datum for the SPR results. After a very simple dehybridization protocol, MALDI-MS analysis allowed detection of the target-ssODNs hybridized on the sensor/array platforms.

中文翻译:

通过使用 SPR、椭偏仪和 MALDI-MS 在传感器/阵列平台上进行杂交

摘要 本研究的目的是开发一种方法,其中表面等离子体共振 (SPR)、椭偏仪 (EM) 和基质辅助激光解吸/电离质谱 (MALDI-MS) 将一起用于检测单链寡脱氧核苷酸 (ssODN) 目标。选定的目标 ssODN 及其互补的探针 ssODN,带有 -SH 端基、间隔臂(HS-(CH2)6–(T)15 和非互补 ssODN)。硅胶基印章带有制备了 16 个区域,用于探针-ssODNs 在镀金表面上的微接触印刷 (μCP)。共固定调节剂-间隔分子(6-巯基-1-己醇)以控制表面探针密度,定位探针-ssODN,并消除非特异性相互作用。SPR 成功地用于跟踪目标 ssODN 与平台表面上固定的探针 ssODN 的杂交。在 100 分钟内实现完全杂交。结果表明,δ 的相对变化与 1 µm 以下的目标浓度之间存在线性关系。使用椭偏仪 (IEM) 的成像版本允许对表面进行成像并支持 SPR 结果的额外数据。在一个非常简单的去杂交协议之后,MALDI-MS 分析允许检测在传感器/阵列平台上杂交的目标 ssODN。使用椭偏仪 (IEM) 的成像版本允许对表面进行成像并支持 SPR 结果的额外数据。在一个非常简单的去杂交协议之后,MALDI-MS 分析允许检测在传感器/阵列平台上杂交的目标 ssODN。使用椭偏仪 (IEM) 的成像版本允许对表面进行成像并支持 SPR 结果的额外数据。在一个非常简单的去杂交协议之后,MALDI-MS 分析允许检测在传感器/阵列平台上杂交的目标 ssODN。
更新日期:2020-05-13
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