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Orientation selection of broad-spectrum aptamers against lipopolysaccharides based on capture-SELEX by using magnetic nanoparticles

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Abstract

The authors describe a new kind of selection method (referred to as orientation selection) for improved screening for broad-spectrum lipopolysaccharides (LPSs) using unlabeled ssDNA aptamers. The method is based on the capture-SELEX technique using magnetic nanoparticles. LPS from Salmonella enterica serotype typhimurium was chosen as an exemplary target. Once the ssDNA library is preconcentrated to a certain degree, two Gram-negative bacteria were used as orientation molecules in the subsequent selection process. Using this strategy, one optimal aptamer ('EA7') was captured that has a high affinity (Kd = 102 ± 17 nM) for LPS and it was also confirmed that it can recognize three other bacterial LPSs. It is presumed that EA7 binds to the lipid A region of LPS. When using carboxyfluorescein labeled EA7, the observed fluorescence intensity and concentrations of four types of LPSs in drinking water are linearly correlated. The lower detection limit of the LPS is 3 ng·mL−1. Compared to multi-target mixed selection and conventional SELEX methods, the new orientation selection strategy produces results that are less uncertain.

An improved orientation selection strategy in capture-SELEX procedures was developed to screen broad-spectrum aptamers against lipopolysaccharides by using magnetic nanoparticles. Two Gram-negative bacteria were used as orientation molecules after library preconcentrated to a certain degree.

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Acknowledgments

This work was supported by National Natural Science Fund of China [21375049], the special fund for innovation in inspection and detection of food and drug of GDFDA [2015ZX07], Key Research and Development Program of Jiangsu Province [BE2017623, BE2016306], the natural science foundation of Jiangsu Province [BK20140155], China Postdoctoral Science Foundation [2016 T90430], and the Technology Research and Development Program of Suzhou [SYN201513].

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Authors and Affiliations

  1. School of Food Science and Technology, Jiangnan University, Wuxi, 214122, China

    Hua Ye, Nuo Duan, Shijia Wu, Huajie Gu & Zhouping Wang

  2. College of Food and Drug, Anhui Science and Technology University, Fengyang, 233100, China

    Hua Ye

  3. Zhongshan Institute for Food and Drug Control, Zhongshan, 528437, China

    Guiliang Tan

  4. China Rural Technology Development Center, Beijing, 100045, China

    Jinghong Li

  5. National Engineering Research Center of Seafood, School of Food Science and Technology, Dalian Polytechnic University, Dalian, 116034, China

    Haitao Wang

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  1. Hua Ye
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  2. Nuo Duan
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Correspondence to Zhouping Wang.

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Ye, H., Duan, N., Wu, S. et al. Orientation selection of broad-spectrum aptamers against lipopolysaccharides based on capture-SELEX by using magnetic nanoparticles. Microchim Acta 184, 4235–4242 (2017). https://doi.org/10.1007/s00604-017-2453-3

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