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Fluorescent aptasensor based on conformational switch–induced hybridization for facile detection of β-amyloid oligomers

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Abstract

Aβ oligomers (AβO) are a dominant biomarker for early Alzheimer’s disease diagnosis. A fluorescent aptasensor coupled with conformational switch–induced hybridization was established to detect AβO. The fluorescent aptasensor is based on the interaction of fluorophore-labeled AβO-specific aptamer (FAM-Apt) against its partly complementary DNA sequence on the surface of magnetic beads (cDNA-MBs). Once the FAM-Apt binds to AβO, the conformational switch of FAM-Apt increases the tendency to be captured by cDNA-MBs. This causes a descending fluorescence of supernatant, which can be utilized to determine the levels of AβO. Thus, the base-pair matching above 12 between FAM-Apt and cDNA-MBs with increasing hybridizing free energies reached the ascending fluorescent signal equilibrium. The optimized aptasensor showed linearity from 1.7 ng mL−1 to 85.1 (R = 0.9977) with good recoveries (79.27–109.17%) in plasma. Furthermore, the established aptasensor possesses rational selectivity in the presence of monomeric Aβ, fibrotic Aβ, and interferences. Therefore, the developed aptasensor is capable of quantifying AβO in human plasma and possesses the potential to apply in clinical cases.

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Abbreviations

AβO:

Amyloid beta peptides

AD:

Alzheimer’s disease

FAM-Apt:

Fluorophore-labeled AβO-specific aptamer

cDNA-MBs:

Complementary DNA sequence on the surface of magnetic beads

CSF:

Cerebrospinal fluid

HFIP:

1,1,1,3,3,3-Hexafluoro-2-propanol

BSA:

Bovine serum albumin

HSA:

Human serum albumin

PTA:

Phosphotungstic acid

SDS:

Sodium dodecyl sulfate

Tris:

Tris(hydroxymethyl)aminomethane

CE:

Capillary electrophoresis

PDA:

Photodiode array

BGE:

Background electrolyte

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Acknowledgements

We gratefully acknowledge the support of the Ministry of Science and Technology of Taiwan (MOST 110-2113-M-194-007).

The application of this developed aptasensor to human plasma acquired from healthy subjects in Kaohsiung Medical University was approved by the Institutional Review Board (IRB) of Kaohsiung Medical University Chung-Ho Memorial Hospital (KMUHIRB-E(I)-20190389).

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Authors and Affiliations

  1. School of Pharmacy, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan

    Chun-Hsien Chen, Chun-Chi Wang & Yen-Ling Chen

  2. Department of Pediatrics, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yuh-Jyh Jong

  3. Translational Research Center of Neuromuscular Diseases, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yuh-Jyh Jong

  4. Department of Laboratory Medicine, Kaohsiung Medical University Hospital, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yuh-Jyh Jong

  5. Graduate Institute of Clinical Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yuh-Jyh Jong

  6. Department of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsinchu, Taiwan

    Yuh-Jyh Jong

  7. Department of Public Health, College of Health Sciences, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yu-Ying Chao

  8. Department of Medical Research, Kaohsiung Medical University Hospital, Kaohsiung, Taiwan

    Chun-Chi Wang

  9. Department of Chemistry and Biochemistry, National Chung Cheng University, Chiayi, Taiwan

    Yen-Ling Chen

  10. Center for Nano Bio-Detection, National Chung Cheng University, Chiayi, Taiwan

    Yen-Ling Chen

  11. Department of Fragrance and Cosmetic Science, College of Pharmacy, Kaohsiung Medical University, Kaohsiung, Taiwan

    Yen-Ling Chen

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  1. Chun-Hsien Chen
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Chen, CH., Jong, YJ., Chao, YY. et al. Fluorescent aptasensor based on conformational switch–induced hybridization for facile detection of β-amyloid oligomers. Anal Bioanal Chem 414, 8155–8165 (2022). https://doi.org/10.1007/s00216-022-04350-7

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