Layer-by-layer generation of PEG-based regenerable immunosensing surfaces for small-sized analytes.
Layer-by-layer generation of PEG-based regenerable immunosensing surfaces for small-sized analytes.
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用于小尺寸分析物的基于 PEG 的可再生免疫传感表面的逐层生成
DOI：
10.1016/j.bios.2014.08.047
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发表时间：
2015
影响因子：
12.6
通讯作者：
Boujday
中科院分区：
文献类型：
--
作者：
Huebner;Ben Haddada;Méthivier;Niessner;Boujday
小分子（半抗原）如药物或肽可以在基于竞争性免疫测定的流通测定中用作抗体结合的靶标。在这项工作中，提出了一种用于制备聚乙二醇（PEG）涂层的策略，用于随后在玻璃型二氧化硅表面上固定半抗原，并详细描述了其特征。使用两个带有末端硅烷醇基团的基底，载玻片和硅晶片。首先，彻底清洁和预处理表面以产生额外的硅烷醇基团。然后，使用3-缩水甘油氧基丙基三甲氧基硅烷（GOPTS）产生具有末端环氧基团的硅烷层。环氧基团用于结合具有末端氨基的二氨基-聚（乙二醇）（DAPEG）层。最后，将低分子量化合物双氯芬酸结合到表面，用作半抗原竞争性生物传感的模型配体。采用原子力显微镜（AFM），水接触角测量，掠角衰减全反射（GA-ATR）FT-IR光谱，和X射线光电子能谱（XPS）的基本步骤，其特征在于。使用这些技术收集的数据已经证实了分子种类的连续接枝，证明了在二氧化硅表面上产生均匀的单层，并验证了所提出的官能化机制。所得表面用于研究多克隆抗双氯芬酸抗体识别和可逆性，使用石英晶体微天平与耗散（QCM-D）测量或自动流通免疫测定与化学发光（CL）读出。对于这两种技术，观察到抗体结合的识别和可逆性。还评估了传感器随时间的稳定性，在水溶液中14天后未观察到CL响应降低。本文提出的表面功能化策略可以在未来用作半抗原生物传感器的可重复使用的通用平台。
Small molecules (haptens) like pharmaceuticals or peptides can serve as targets for antibody binding in competitive immunoassay-based flow-through assays. In this work, a strategy for preparing polyethylene glycol (PEG) coatings for subsequent hapten immobilization on glass-type silica surfaces is presented and characterized in detail. Two substrates bearing terminal silanol groups were utilized, a glass slide and a silicon wafer. First, surfaces were thoroughly cleaned and pretreated to generate additional silanol groups. Then, a silane layer with terminal epoxy groups was created using 3-glycidyloxypropyltrimethoxysilane (GOPTS). Epoxy groups were used to bind a layer of diamino-poly(ethylene glycol) (DAPEG) with terminal amino groups. Finally, the low molecular weight compound diclofenac was bound to the surface to be used as model ligand for competitive biosensing of haptens. The elementary steps were characterized using atomic force microscopy (AFM), water contact angle measurement, grazing-angle attenuated total reflection (GA-ATR) FT-IR spectroscopy, and X-ray photoelectron spectroscopy (XPS). The data collected using these techniques have confirmed the successive grafting of the molecular species, evidencing, that homogeneous monolayers were created on the silica surfaces and validated the proposed mechanism of functionalization. The resulting surfaces were used to investigate polyclonal anti-diclofenac antibodies recognition and reversibility using quartz crystal microbalance with dissipation (QCM-D) measurements or an automated flow-through immunoassay with chemiluminescence (CL) read-out. For both techniques, recognition and reversibility of the antibody binding were observed. The stability of sensors over time was also assessed and no decrease in CL response was observed upon 14 days in aqueous solution. The herein presented strategy for surface functionalization can be used in the future as reproducible and reusable universal platform for hapten biosensors.
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影响因子：
4.3
作者：
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通讯作者：
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影响因子：
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通讯作者：
J. Pillot
DOI：
--
发表时间：
2000
期刊：
影响因子：
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作者：
X. Su;F. Chew;Sam F. Y. Li
通讯作者：
Sam F. Y. Li
影响因子：
3.9
作者：
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影响因子：
3.9
作者：
T. Lummerstorfer;H. Hoffmann
通讯作者：
H. Hoffmann