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A facile aptamer immobilization strategy to fabricate a robust affinity monolith for highly specific in-tube solid-phase microextraction
Analyst ( IF 3.6 ) Pub Date : 2021-08-11 , DOI: 10.1039/d1an00993a Tingting Zhao 1 , Shiqian Tong 1 , Susu Zhou 1 , Chenchen Lin 1 , Xucong Lin 1, 2 , Zenghong Xie 1, 2
Analyst ( IF 3.6 ) Pub Date : 2021-08-11 , DOI: 10.1039/d1an00993a Tingting Zhao 1 , Shiqian Tong 1 , Susu Zhou 1 , Chenchen Lin 1 , Xucong Lin 1, 2 , Zenghong Xie 1, 2
Affiliation
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Developing a functional affinity monolithic column towards in-tube solid-phase microextraction (IT-SPME) for selective sample pretreatment is critical. Herein, a high-performance capillary affinity monolithic column with an ultra-high aptamer coverage density was rapidly fabricated via a simple adsorption strategy, in which aptamers with natural sequences were directly immobilized on an ammonium-based strongly cationic matrix. Limitations of the traditional biological or covalent methods such as time-consuming modification reactions, special requirement of active groups (e.g. –NH2 and –SH) on the aptamer, and low aptamer coverage density levels were avoided. An ultra-high coverage density of 8616 pmol μL−1 was achieved with excellent stability, and the highest aptamer-modification level among all the current methods was reached. Selective recognition and high recovery yields of the model mycotoxin ochratoxin A (OTA) were achieved in 95.9 ± 0.98%–97.9 ± 0.28% (n = 3). In particular, there was little cross-reactivity towards the OTB analogue of only 0.5% even in the case of 250 fold of the analogue OTB, which was not reported in previous affinity monoliths. Upon sample analysis, satisfactory discriminations of trace OTA were obtained at 93.7 ± 1.4%–95.5 ± 2.5% (n = 3) in beer and wheat. A facile and direct method for efficiently fabricating an aptamer-based affinity monolith towards online selective IT-SPME was proposed.
中文翻译：
一种简便的适体固定策略，用于制造用于高度特异性管内固相微萃取的强大亲和整体
开发用于选择性样品预处理的管内固相微萃取 (IT-SPME) 的功能亲和整体柱是至关重要的。在此，通过简单的吸附策略快速制造了具有超高适配体覆盖密度的高性能毛细管亲和整体柱，其中具有天然序列的适配体直接固定在铵基强阳离子基质上。避免了传统生物学或共价方法的局限性，例如耗时的修饰反应、适体对活性基团（例如-NH 2和-SH）的特殊要求以及适体覆盖密度水平低等。8616 pmol μL -1的超高覆盖密度以优异的稳定性实现，并且达到了目前所有方法中最高的适配体修饰水平。模型赭曲霉毒素 A (OTA) 的选择性识别和高回收率达到了 95.9 ± 0.98%–97.9 ± 0.28% ( n = 3)。特别是，即使在类似物 OTB 的 250 倍的情况下，对 OTB 类似物的交叉反应性也很少，只有 0.5%，这在以前的亲和整体中没有报道。根据样品分析，在啤酒和小麦中获得了 93.7 ± 1.4%–95.5 ± 2.5% ( n = 3)的痕量 OTA 的令人满意的区分。提出了一种简便而直接的方法，用于有效地制造基于适体的亲和单体，用于在线选择性 IT-SPME。
更新日期：2021-08-25
中文翻译：
一种简便的适体固定策略，用于制造用于高度特异性管内固相微萃取的强大亲和整体
开发用于选择性样品预处理的管内固相微萃取 (IT-SPME) 的功能亲和整体柱是至关重要的。在此，通过简单的吸附策略快速制造了具有超高适配体覆盖密度的高性能毛细管亲和整体柱，其中具有天然序列的适配体直接固定在铵基强阳离子基质上。避免了传统生物学或共价方法的局限性，例如耗时的修饰反应、适体对活性基团（例如-NH 2和-SH）的特殊要求以及适体覆盖密度水平低等。8616 pmol μL -1的超高覆盖密度以优异的稳定性实现，并且达到了目前所有方法中最高的适配体修饰水平。模型赭曲霉毒素 A (OTA) 的选择性识别和高回收率达到了 95.9 ± 0.98%–97.9 ± 0.28% ( n = 3)。特别是，即使在类似物 OTB 的 250 倍的情况下，对 OTB 类似物的交叉反应性也很少，只有 0.5%，这在以前的亲和整体中没有报道。根据样品分析，在啤酒和小麦中获得了 93.7 ± 1.4%–95.5 ± 2.5% ( n = 3)的痕量 OTA 的令人满意的区分。提出了一种简便而直接的方法，用于有效地制造基于适体的亲和单体，用于在线选择性 IT-SPME。
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