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Insights into the Ochratoxin A/Aptamer Interactions on a Functionalized Silicon Surface by Fourier Transform Infrared and UV–Vis Studies
Langmuir ( IF 3.7 ) Pub Date : 2020-11-09 , DOI: 10.1021/acs.langmuir.0c02358 Timothy Aschl 1 , Gilles Frison 2, 3 , Anne Moraillon 1 , François Ozanam 1 , Philippe Allongue 1 , Anne Chantal Gouget-Laemmel 1
Langmuir ( IF 3.7 ) Pub Date : 2020-11-09 , DOI: 10.1021/acs.langmuir.0c02358 Timothy Aschl 1 , Gilles Frison 2, 3 , Anne Moraillon 1 , François Ozanam 1 , Philippe Allongue 1 , Anne Chantal Gouget-Laemmel 1
Affiliation
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The association of a mycotoxin—ochratoxin A (OTA)—with a high-affinity DNA aptamer (anti-OTA) immobilized on a functionalized surface has been investigated at the molecular level. Anti-OTA aptamers are coupled by aminolysis in several steps on an acid-terminated alkyl monolayer grafted on a silicon substrate, and Fourier transform infrared spectroscopy in attenuated total reflection geometry is used to assess the immobilization of anti-OTA (in its unfolded single-strand form) and determine its areal density (ca. 1.4/nm2). IR spectra further demonstrate that the OTA/anti-OTA association is efficient and selective and that several association/dissociation cycles may be conducted on the same surface. The areal density of OTA measured after association on the surface (IR spectroscopy) and after dissociation from the surface (UV–vis spectroscopy) falls in the range 0.16–0.3/nm2 which is close to the areal density of a closed-packed monolayer of anti-OTA aptamers folded to form their G-quadruplex structure. The interactions between OTA and its aptamer at the surface are discussed with the help of density functional theory calculations—to identify the complex IR vibrational modes of OTA in solution—and UV–vis spectroscopy—to determine the protonation state of the adsorbing species (i.e., OTA dissolved in the buffer solution).
中文翻译：
通过傅立叶变换红外和紫外-可见光谱研究洞察功能化硅表面上O曲毒素A /适体的相互作用
在分子水平上已经研究了霉菌毒素—曲霉毒素A（OTA）与固定在功能化表面上的高亲和力DNA适体（anti-OTA）的关联。抗OTA适体通过在接枝到硅基板上的酸封端的烷基单分子层上的氨解反应分几步进行，并且使用傅立叶变换红外光谱在衰减的全反射几何结构中评估抗OTA的固定化（在其展开的单分子-线形）并确定其面密度（约1.4 / nm 2）。红外光谱进一步证明OTA /抗OTA缔合是有效和选择性的，并且可以在同一表面上进行多个缔合/解离循环。在表面缔合后（红外光谱）和从表面解离（紫外可见光谱）后测得的OTA面密度在0.16-0.3 / nm 2范围内，接近密闭单层膜的面密度的抗-OTA适体折叠形成其G-四链体结构。借助密度泛函理论计算（以确定溶液中OTA的复杂IR振动模式）和UV-可见光谱，讨论了OTA及其表面适体之间的相互作用，并确定了吸附物质的质子化状态（即，将OTA溶解在缓冲溶液中）。
更新日期：2020-11-25
中文翻译：
通过傅立叶变换红外和紫外-可见光谱研究洞察功能化硅表面上O曲毒素A /适体的相互作用
在分子水平上已经研究了霉菌毒素—曲霉毒素A（OTA）与固定在功能化表面上的高亲和力DNA适体（anti-OTA）的关联。抗OTA适体通过在接枝到硅基板上的酸封端的烷基单分子层上的氨解反应分几步进行，并且使用傅立叶变换红外光谱在衰减的全反射几何结构中评估抗OTA的固定化（在其展开的单分子-线形）并确定其面密度（约1.4 / nm 2）。红外光谱进一步证明OTA /抗OTA缔合是有效和选择性的，并且可以在同一表面上进行多个缔合/解离循环。在表面缔合后（红外光谱）和从表面解离（紫外可见光谱）后测得的OTA面密度在0.16-0.3 / nm 2范围内，接近密闭单层膜的面密度的抗-OTA适体折叠形成其G-四链体结构。借助密度泛函理论计算（以确定溶液中OTA的复杂IR振动模式）和UV-可见光谱，讨论了OTA及其表面适体之间的相互作用，并确定了吸附物质的质子化状态（即，将OTA溶解在缓冲溶液中）。
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