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Yujing Ding, Zihan Gao, Hao Li,
Real milk sample assisted selection of specific aptamer towards sarafloxacin and its application in establishment of an effective aptasensor,
Sensors and Actuators B: Chemical,
Volume 343,
2021,
130113,
ISSN 0925-4005,
https://doi.org/10.1016/j.snb.2021.130113.
(http://www.sciencedirect.com.group21-s.aronip.com/science/article/pii/S0925400521006821)
Abstract: The aptamers screened by the classic SELEX strategy or other approaches do not undergo the real sample analysis during the selection process, and the environmental inconsistency between the selection and final detection systems might influence the aptamer characteristics and detection performance. Herein, a novel real sample assisted SELEX selection strategy has been proposed to rationally select the suitable aptamers with high detection performance, using the selection of a sarafloxacin  specific aptamer as an example. Based on the developed strategy, the aptamer Apt-F2 with higher specificity and SAR binding affinity was obtained as compared to the aptamer Apt-B4 selected by the standard SELEX strategy. Apt-F2 also exhibited a stronger conformational change after binding as compared to Apt-B4. Apt-B4 and Apt-F2 were used as a platform for developing the fluorescent aptasensor. Under optimized conditions, the limit of detection of Apt-B4- and Apt-F2-based aptasensors for SAR was 0.19 and 0.12 ng/mL respectively. Moreover, Apt-F2-based aptasensor also revealed a slightly superior SAR recovery rate from the spiked milk samples. The findings obtained in the study confirmed the advantage of the real SELEX strategy as compared to the traditional version. Further, the characteristics of the aptasensor proposed here also revealed its application potential for detecting SAR in the real milk or other animal-sourced food samples.
Keywords: Real SELEX strategy; Aptamer selection; Sarafloxacin; Milk; Aptasensor

摘要：通过经典的SELEX策略或其他方法筛选出的适体在筛选过程中没有经过真实的样本分析，而且筛选系统与最终检测系统之间的环境不一致可能影响适体的特性和检测性能。本文以萨拉氟沙星特异性核酸适体的选择为例，提出了一种新的真实样本辅助SELEX选择策略，以合理选择具有高检测性能的适体。基于该策略，获得了比标准SELEX策略选择的Apt-B4具有更高特异性和SAR结合亲和力的Apt-F2适体。与Apt-B4相比，Apt-F2在结合后表现出更强的构象变化。以Apt-B4和Apt-F2为平台，研制了荧光型aptasensor。在优化的条件下，Apt-B4-和Apt-F2-基aptasensors对SAR的检测限分别为0.19和0.12 ng/mL。此外，基于Apt-F2的aptasensor还显示，从加标牛奶样品中获得的SAR回收率略高。研究结果证实了真实SELEX策略相对于传统策略的优势。此外，本文提出的aptasensor的特性也揭示了其在实际牛奶或其他动物源食品样品中检测SAR的应用潜力。

关键词：实SELEX策略；适体选择；沙拉沙星；牛奶；适体传感器

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