title: Gonyautoxin 1/4 aptamers with high-affinity and high-specificity: From efficient selection to aptasensor application.
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.
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China.
authors.forename: Shunxiang
authors.initials: S
authors.lastname: Gao
authors.affiliation: 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.
authors.forename: Bo
authors.initials: B
authors.lastname: Hu
authors.affiliation: 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.
authors.forename: Xin
authors.initials: X
authors.lastname: Zheng
authors.affiliation: Beijing Institute of Pharmaceutical Chemistry, Beijing 102205, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Cao
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China.
authors.forename: Dejing
authors.initials: D
authors.lastname: Liu
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China.
authors.forename: Mingjuan
authors.initials: M
authors.lastname: Sun
authors.affiliation: 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.
authors.forename: Binghua
authors.initials: B
authors.lastname: Jiao
authors.affiliation: Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China. Electronic address: lhwang@smmu.edu.cn.
authors.forename: Lianghua
authors.initials: L
authors.lastname: Wang
mesh_terms: D000818:Animals; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D057230:Limit of Detection; D008387:Marine Toxins / Q000032:analysis*; D015203:Reproducibility of Results; D052156:SELEX Aptamer Technique; D012530:Saxitoxin / Q000031:analogs & derivatives* / Q000032:analysis; D012758:Shellfish / Q000032:analysis*
chemical_list: D052157:Aptamers, Nucleotide; D008387:Marine Toxins; D012530:Saxitoxin; C013007:gonyautoxins
keywords: Aptasensor; Biolayer interferometry; In vitro selection; Optimization