2. Academic Validation
  3. Gonyautoxin 1/4 aptamers with high-affinity and high-specificity: From efficient selection to aptasensor application

Gonyautoxin 1/4 aptamers with high-affinity and high-specificity: From efficient selection to aptasensor application

  • Biosens Bioelectron. 2016 May 15:79:938-44. doi: 10.1016/j.bios.2016.01.032.
Shunxiang Gao 1,  Bo Hu 2,  Xin Zheng 3,  Ying Cao 4,  Dejing Liu 1,  Mingjuan Sun 1,  Binghua Jiao 5,  Lianghua Wang 6
Affiliations

Affiliations

  • 1 Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China.
  • 2 Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China; Marine Biological Institute, College of Marine Military Medicine, Second Military Medical University, Shanghai 200433, China.
  • 3 Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China; Department of Laboratory Diagnosis, Changhai hospital, Second Military Medical University, Shanghai 200433, China.
  • 4 Beijing Institute of Pharmaceutical Chemistry, Beijing 102205, China.
  • 5 Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China; Marine Biological Institute, College of Marine Military Medicine, Second Military Medical University, Shanghai 200433, China. Electronic address: bhjiao@smmu.edu.cn.
  • 6 Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China. Electronic address: lhwang@smmu.edu.cn.
PMID: 26802576 DOI: 10.1016/j.bios.2016.01.032
Abstract

Gonyautoxin 1/4 (GTX1/4) are potent marine neurotoxins with significant public health impact. However, the ethical issues and technical defects associated with the currently applied detection methods for paralytic shellfish toxin GTX1/4 are pressing further studies to develop suitable alternatives in a regulatory monitoring system. This work describes the first successful selection, optimization, and characterization of an aptamer that bind with high affinity and specificity to GTX1/4. Compared to the typical MB-SELEX, GO-SELEX, an advanced screening technology, has significant advantages for small molecular aptamer development. Furthermore, we truncated GTX1/4 aptamer and obtained the aptamer core sequence with a higher Kd of 17.7 nM. The aptamer GO18-T-d was then used to construct a label-free and real-time optical BLI aptasensor for the detection of GTX1/4. The aptasensor showed a broad detection range from 0.2 to 200 ng/mL GTX1/4 (linear range from 0.2 to 90 ng/mL), with a low detection limit of 50 pg/mL. Moreover, the aptasensor exhibited a high degree of specificity for GTX1/4 and no cross reactivity to Other marine toxins. The aptasensor was then applied to the detection of GTX1/4 in spiked shellfish samples and showed a good reproducibility and stability. We believe that this novel aptasensor offers a promising alternative to traditional analytical methods for the rapid detection of the marine biotoxin GTX1/4.

Keywords

Aptasensor; Biolayer interferometry; In vitro selection; Optimization.

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