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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：通過經典的SELEX策略或其他方法篩選出的適體在篩選過程中沒有經過真實的樣本分析，而且篩選系統與最終檢測系統之間的環境不一致可能影響適體的特性和檢測性能。本文以薩拉氟沙星特異性核酸適體的選擇為例，提出了一種新的真實樣本輔助SELEX選擇策略，以合理選擇具有高檢測性能的適體?；谠摬呗?，獲得了比標準SELEX策略選擇的Apt-B4具有更高特異性和SAR結合親和力的Apt-F2適體。與Apt-B4相比，Apt-F2在結合后表現出更強的構象變化。以Apt-B4和Apt-F2為平臺，研制了熒光型aptasensor。在優化的條件下，Apt-B4-和Apt-F2-基aptasensors對SAR的檢測限分別為0.19和0.12?ng/mL。此外，基于Apt-F2的aptasensor還顯示，從加標牛奶樣品中獲得的SAR回收率略高。研究結果證實了真實SELEX策略相對于傳統策略的優勢。此外，本文提出的aptasensor的特性也揭示了其在實際牛奶或其他動物源食品樣品中檢測SAR的應用潛力。

關鍵詞：實SELEX策略；適體選擇；沙拉沙星；牛奶；適體傳感器

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