Self-assembly strategy to reduce non-specific adsorption for the development of high sensitivity quantitative immunoassay
Self-assembly strategy to reduce non-specific adsorption for the development of high sensitivity quantitative immunoassay
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减少非特异性吸附的自组装策略，用于开发高灵敏度定量免疫分析
DOI：
10.1016/j.aca.2022.340367
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发表时间：
2022
影响因子：
6.2
通讯作者：
Lin Song Li
中科院分区：
文献类型：
--
作者：
Yanbing Lv;Man Zhao;Jinjin Fan;Ruili Wu;Yanxia Xu;Jinjie Li;Ning Li;Huaibin Shen;Fang Guo;Lin Song Li
开发具有低非特异性吸附的功能化表面对于其生物医学应用非常重要。为了抑制玻璃基板上的非特异性吸附，我们通过自组装在基板表面修饰一层致密的带负电的薄膜（SO 3 2− ），设计了一种新型光学生物芯片。与未处理的玻璃基板相比，聚(苯乙烯磺酸)钠盐(PSS)或内消旋四(4-磺基苯基)卟啉二盐酸盐(TSPP)在单独的改性玻璃基板上的吸附减少了约300倍或400倍。考虑到混合后TSPP与溶液中量子点之间的荧光共振能量转移（FRET）效应，设计了2层TSPP后4层PSS的策略来改性制备生物芯片的玻璃。在优化条件下，TSPP和PSS共处理的功能化玻璃基板上的生物芯片实现了基于量子点荧光免疫吸附测定（QD-FLISA）的C反应蛋白（CRP）的灵敏定量检测。使用 TSPP 和 PSS 共处理的玻璃基板表面，CRP 的检测限 (LOD) 达到 0.69 ng/mL，范围为 1-1000 ng/mL，对 PSS 修饰的生物芯片和 TSPP 修饰的生物芯片的灵敏度分别提高约 1.9 倍和 7.5 倍。这项工作展示了一种有效且便捷的策略，可以在功能化表面上获得具有低非特异性吸附特性的生物芯片，从而为在玻璃基板上创建超高灵敏度微通道或微阵列提供新方法。 • 研究了玻璃基板上非特异性吸附的原因。 • 制备并评估致密的带负电的聚合物基底(SO 3 2− )。 • 在经TSPP 和PSS 共同处理的功能化玻璃基板上构建了一种新型光学生物芯片，以抑制非特异性吸附。 • 所设计的生物芯片能够实现基于QD-FLISA的CRP的灵敏定量检测。
The development of functionalized surfaces with low non-specific adsorption is important for their biomedical applications. To inhibit non-specific adsorption on glass substrate, we designed a novel optical biochip by modifying a layer of dense negatively charged film (SO 3 2− ) on its substrate surface via self-assembly. Compared with the untreated glass substrate, it reduced the adsorption by about 300-fold or 400-fold by poly (styrene sulfonic acid) sodium salt (PSS), or meso -tetra (4-sulfonatophenyl) porphine dihydrochloride (TSPP) on individually the modified glass substrate. Considering the effect of fluorescence resonance energy transfer (FRET) between TSPP and the QDs in solution by mixing, a strategy of 2-layer of TSPP followed by 4-layer of PSS was designed to modify the glass for preparing biochips. Under the optimized conditions, the biochip on functionalized glass substrates co-treated with TSPP and PSS realized the sensitive quantitative detection of C-reactive protein (CRP) based on a quantum dot fluorescence immunosorbent assay (QD-FLISA). The limit of detection (LOD) for CRP achieved 0.69 ng/mL with the range of 1-1000 ng/mL using TSPP and PSS co-treated glass substrate surfaces, which was respectively about 1.9-fold and 7.5-fold more sensitive to the PSS-modified biochip and the TSPP-modified biochip. This work demonstrated an effective and convenient strategy to obtain biochips with low non-specific adsorption properties on functionalized surfaces, thus providing a new approach for creating ultra-high sensitivity microchannels or microarrays on glass substrates. • The causes of non-specific adsorption on glass substrates were studied. • The dense negatively charged polymer substrates (SO 3 2− ) were prepared and evaluated. • A novel optical biochip on functionalized glass substrates co-treated with TSPP and PSS was constructed to inhibit non-specific adsorption. • The designed biochip can realize the sensitive quantitative detection of CRP based on QD-FLISA.