Enzyme-linked, aptamer-based, competitive biolayer interferometry biosensor for palytoxin.
In this study, we coupled biolayer interferometry (BLI) with competitive binding assay through an enzyme-linked aptamer and developed a real-time, ultra-sensitive, rapid quantitative method for detection of the marine biotoxin palytoxin. Horseradish peroxidase-labeled aptamers were used as biorecognition receptors to competitively bind with palytoxin, which was immobilized on the biosensor surface. The palytoxin: horseradish peroxidase-aptamer complex was then submerged in a 3,3'-diaminobenzidine solution, which resulted in formation of a precipitated polymeric product directly on the biosensor surface and a large change in the optical thickness of the biosensor layer. This change could obviously shift the interference pattern and generate a response profile on the BLI biosensor. The biosensor showed a broad linear range for palytoxin (200-700pg/mL) with a low detection limit (0.04pg/mL). Moreover, the biosensor was applied to the detection of palytoxin in spiked extracts and showed a high degree of selectivity for palytoxin, good reproducibility, and stability. This enzyme-linked, aptamer-based, competitive BLI biosensor offers a promising method for rapid and sensitive detection of palytoxin and other analytes.
Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China; Research Center, Eye & ENT Hospital, Fudan University, Shanghai, 200032 China; Shanghai Key Laboratory of Visual Impairment and Restoration, Shanghai 200032, China; Key Laboratory of Myopia, Ministry of Health, Fudan University, Shanghai 200032, China.
Shunxiang
S
Gao
Department of Laboratory Diagnosis, Changhai Hospital, Second Military Medical University, Shanghai 200433, China.
Xin
X
Zheng
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.
Bo
B
Hu
Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China.
Mingjuan
M
Sun
Research Center, Eye & ENT Hospital, Fudan University, Shanghai, 200032 China; Shanghai Key Laboratory of Visual Impairment and Restoration, Shanghai 200032, China; Key Laboratory of Myopia, Ministry of Health, Fudan University, Shanghai 200032, China. Electronic address: jihongwu@fudan.edu.cn.
Jihong
J
Wu
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.
Binghua
B
Jiao
Department of Biochemistry and Molecular Biology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China. Electronic address: lhwang@smmu.edu.cn.
Lianghua
L
Wang
D015100:3,3'-Diaminobenzidine / Q000737:chemistry; D000178:Acrylamides / Q000737:chemistry / Q000302:isolation & purification*; D052157:Aptamers, Nucleotide / Q000737:chemistry*; D015374:Biosensing Techniques; D003064:Cnidarian Venoms; D006735:Horseradish Peroxidase / Q000737:chemistry; D007368:Interferometry; D057230:Limit of Detection; D000095702:Polyether Toxins
D015100:3,3'-Diaminobenzidine; D000178:Acrylamides; D052157:Aptamers, Nucleotide; D003064:Cnidarian Venoms; D006735:Horseradish Peroxidase; C010272:palytoxin; D000095702:Polyether Toxins
Aptamer; Biolayer interferometry; Biosensor; Palytoxin; SELEX