An aptamer based fluorometric assay for amyloid-β oligomers using a metal-organic framework of type Ru@MIL-101(Al) and enzyme-assisted recycling


Amyloid-beta (Aβ) oligomers causing neuron damage are regarded as potential therapeutic targets and diagnostic markers for Alzheimer’s disease (AD). A homogeneous turn-on fluorometric aptasensor is described for Aβ oligomers. It is highly selective and non-invasive and based on (a) the use of a luminescent metal-organic framework carrying aptamer-modified AuNPs (L-MOF/Apt-Au) as tracking agent, and (b) enzyme-assisted target recycling signal amplification. The tracking agent does not emit fluoresce by fluorescence resonance energy transfer (FRET) between the luminescent MOF as donor and Apt-Au as the acceptor under the excitation wavelength of 466 nm. When Aβ oligomers are added to the tracking agent solution, the Apt-Au on tracking agent can preferentially bind with Aβ oligomers and then be released. This turns the “off” signal of the luminescent MOF tracer to the “on” state. The enzyme (Rec Jf exonuclease) added into the supernatant further improves sensitivity due to enzyme-assisted target-recycling signal amplification. The assay has an excellent linear response to Aβ oligomers from 1.0 pM to 10 nM, with a detection limit of 0.3 pM. This homogeneous turn-on fluorometric method is expected to have potential and applications in clinical diagnosis.

Schematic representation of fluorometric assay for amyloid-β oligomers based on luminescence metal-organic framework nanocomposites as tracking agent with exonuclease-assisted target recycling.

This is a preview of subscription content, access via your institution.

We’re sorry, something doesn't seem to be working properly.

Please try refreshing the page. If that doesn't work, please contact support so we can address the problem.

Scheme 1
Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5


  1. 1.

    Qin J, Park JS, Jo DG, Cho M, Lee Y (2018) Curcumin-based electrochemical sensor of amyloid-β oligomer for the early detection of Alzheimer’s disease. Sensors Actuators B Chem 273:1593–1599.

    CAS  Article  Google Scholar 

  2. 2.

    Chae MS, Yoo YK, Kim J, Kim TG, Hwang KS (2018) Graphene-based enzyme-modified field-effect transistor biosensor for monitoring drug effects in Alzheimer’s disease treatment. Sensors Actuators B Chem 272:448–458.

    CAS  Article  Google Scholar 

  3. 3.

    Ding S, Cao S, Liu Y, Lian Y, Zhu A, Shi G (2017) Rational design of a stimuli-responsive polymer electrode interface coupled with in vivo microdialysis for measurement of sialic acid in live mouse brain in Alzheimer’s disease. ACS Sensors 2(3):394–400.

    CAS  Article  Google Scholar 

  4. 4.

    Feng Y, Wang XP, Yang SG, Wang YJ, Zhang X, Du XT, Liu RT (2009) Resveratrol inhibits beta-amyloid oligomeric cytotoxicity but does not prevent oligomer formation. Neurotoxicology 30(6):986–995.

    CAS  Article  Google Scholar 

  5. 5.

    Lee EB, Leng LZ, Zhang B, Kwong L, Trojanowski JQ, Abel T, Lee VMY (2006) Targeting amyloid-β peptide (Aβ) oligomers by passive immunization with a conformation-selective monoclonal antibody improves learning and memory in Aβ precursor protein (APP) transgenic mice. J Biol Chem 281(7):4292–4299.

    CAS  Article  Google Scholar 

  6. 6.

    Salvadores N, Shahnawaz M, Scarpini E, Tagliavini F, Soto C (2014) Detection of misfolded Aβ oligomers for sensitive biochemical diagnosis of Alzheimer’s disease. Cell Rep 7(1):261–268.

    CAS  Article  Google Scholar 

  7. 7.

    El-Agnaf OM, Salem SA, Paleologou KE, Curran MD, Gibson MJ, Court JA, Allsop D (2006) Detection of oligomeric forms of α-synuclein protein in human plasma as a potential biomarker for Parkinson’s disease. FASEB J 20(3):419–425.

    CAS  Article  Google Scholar 

  8. 8.

    Yu Y, Sun X, Tang D, Li C, Zhang L, Nie D, Shi G (2015) Gelsolin bound β-amyloid peptides (1–40/1–42): electrochemical evaluation of levels of soluble peptide associated with Alzheimer's disease. Biosens Bioelectron 68:115–121.

    CAS  Article  Google Scholar 

  9. 9.

    Jayasena SD (1999) Aptamers: an emerging class of molecules that rival antibodies in diagnostics. Clin Chem 45(9):1628–1650

    CAS  Article  Google Scholar 

  10. 10.

    Chen A, Yang S (2015) Replacing antibodies with aptamers in lateral flow immunoassay. Biosens Bioelectron 71:230–242.

    CAS  Article  Google Scholar 

  11. 11.

    Zhu L, Zhang J, Wang F, Wang Y, Lu L, Feng C, Zhang W (2016) Selective amyloid β oligomer assay based on abasic site-containing molecular beacon and enzyme-free amplification. Biosens Bioelectron 78:206–212.

    CAS  Article  Google Scholar 

  12. 12.

    Hughes ZE, Walsh TR (2017) Structural disruption of an adenosine-binding DNA aptamer on graphene: implications for Aptasensor design. ACS Sensors 2(11):1602–1611.

    CAS  Article  Google Scholar 

  13. 13.

    Liu S, Cheng C, Liu T, Wang L, Gong H, Li F (2015) Highly sensitive fluorescence detection of target DNA by coupling exonuclease-assisted cascade target recycling and DNAzyme amplification. Biosens Bioelectron 63:99–104.

    CAS  Article  Google Scholar 

  14. 14.

    Miao Y, Gan N, Li T, Cao Y, Hu F, Chen Y (2016) An ultrasensitive fluorescence aptasensor for chloramphenicol based on FRET between quantum dots as donor and the magnetic SiO2@ au NPs probe as acceptor with exonuclease-assisted target recycling. Sensors Actuators B Chem 222:1066–1072.

    CAS  Article  Google Scholar 

  15. 15.

    Huang YL, Gao ZF, Luo HQ, Li NB (2017) Sensitive detection of HIV gene by coupling exonuclease III-assisted target recycling and guanine nanowire amplification. Sensors Actuators B Chem 238:1017–1023.

    CAS  Article  Google Scholar 

  16. 16.

    Pham TT, Yin J, Eid JS, Adams E, Lam R, Turner SW, Hanes JW (2016) Single-locus enrichment without amplification for sequencing and direct detection of epigenetic modifications. Mol Gen Genomics 291(3):1491–1504.

    CAS  Article  Google Scholar 

  17. 17.

    Chen M, Gan N, Zhou Y, Li T, Xu Q, Cao Y, Chen Y (2016) An electrochemical aptasensor for multiplex antibiotics detection based on metal ions doped nanoscale MOFs as signal tracers and RecJf exonuclease-assisted targets recycling amplification. Talanta 161:867–874.

    CAS  Article  Google Scholar 

  18. 18.

    Zhu X, Su Q, Feng W, Li F (2017) Anti-stokes shift luminescent materials for bio-applications. Chem Soc Rev 46(4):1025–1039.

    CAS  Article  Google Scholar 

  19. 19.

    Fan C, Lv X, Liu F, Feng L, Liu M, Cai Y, Wang H (2018) Silver nanoclusters encapsulated into metal–organic frameworks with enhanced fluorescence and specific ion accumulation toward the microdot array-based fluorimetric analysis of copper in blood. ACS Sensors 3(2):441–450.

    CAS  Article  Google Scholar 

  20. 20.

    Zhang Y, Yuan S, Day G, Wang X, Yang X, Zhou HC (2018) Luminescent sensors based on metal-organic frameworks. Coord Chem Rev 354:28–45.

    CAS  Article  Google Scholar 

  21. 21.

    Zhang L, Kang Z, Xin X, Sun D (2016) Metal–organic frameworks based luminescent materials for nitroaromatics sensing. CrystEngComm 18(2):193–206.

    CAS  Article  Google Scholar 

  22. 22.

    Chen S, Shi Z, Qin L, Jia H, Zheng H (2016) Two new luminescent cd (II)-metal–organic frameworks as bifunctional chemosensors for detection of cations Fe3+, anions CrO4 2−, and Cr2O7 2−in aqueous solution. Cryst Growth Des 17(1):67–72.

    CAS  Article  Google Scholar 

  23. 23.

    Lian X, Fang Y, Joseph E, Wang Q, Li J, Banerjee S, Zhou HC (2017) Enzyme–MOF (metal–organic framework) composites. Chem Soc Rev 46(11):3386–3401.

    CAS  Article  Google Scholar 

  24. 24.

    Fang JM, Leng F, Zhao XJ, Hu XL, Li YF (2014) Metal–organic framework MIL-101 as a low background signal platform for label-free DNA detection. Analyst 139(4):801–806.

    CAS  Article  Google Scholar 

  25. 25.

    Maza WA, Haring AJ, Ahrenholtz SR, Epley CC, Lin SY, Morris AJ (2016) Ruthenium (II)-polypyridyl zirconium (IV) metal–organic frameworks as a new class of sensitized solar cells. Chem Sci 7(1):719–727.

    CAS  Article  Google Scholar 

  26. 26.

    Radenković S, Antić M, Savić ND, Glišić BĐ (2017) The nature of the au–N bond in gold (iii) complexes with aromatic nitrogen-containing heterocycles: the influence of au (iii) ions on the ligand aromaticity. New J Chem 41(21):12407–12415.

    Article  Google Scholar 

  27. 27.

    Yin HQ, Yang JC, Yin XB (2017) Ratiometric fluorescence sensing and real-time detection of water in organic solvents with one-pot synthesis of Ru@ MIL-101 (Al)–NH2. Anal Chem 89(24):13434–13440.

    CAS  Article  Google Scholar 

  28. 28.

    Chołuj A, Zieliński A, Grela K, Chmielewski MJ (2016) Metathesis@ MOF: simple and robust immobilization of olefin metathesis catalysts inside (Al) MIL-101-NH2. ACS Catal 6(10):6343–6349.

    CAS  Article  Google Scholar 

  29. 29.

    El-Mehalmey WA, Ibrahim AH, Abugable AA, Hassan MH, Haikal RR, Karakalos SG, Alkordi MH (2018) Metal–organic framework@ silica as a stationary phase sorbent for rapid and cost-effective removal of hexavalent chromium. J Mater Chem A 6(6):2742–2751.

    CAS  Article  Google Scholar 

  30. 30.

    Lapitan LD Jr, Guo Y, Zhou D (2015) Nano-enabled bioanalytical approaches to ultrasensitive detection of low abundance single nucleotide polymorphisms. Analyst 140(12):3872–3887.

    CAS  Article  Google Scholar 

  31. 31.

    Lv G, Sun A, Wei P, Zhang N, Lan H, Yi T (2016) A spiropyran-based fluorescent probe for the specific detection of β-amyloid peptide oligomers in Alzheimer’s disease. Chem Commun 52(57):8865–8868.

    CAS  Article  Google Scholar 

  32. 32.

    Park MC, Kim M, Lim GT, Kang SM, An SSA, Kim TS, Kang JY (2016) Droplet-based magnetic bead immunoassay using microchannel-connected multiwell plates (μCHAMPs) for the detection of amyloid beta oligomers. Lab Chip 16(12):2245–2253.

    CAS  Article  Google Scholar 

  33. 33.

    Zhang X, Liu S, Song X, Wang H, Wang J, Wang Y, Huang J, Yu J (2019) Robust and universal SERS sensing platform for multiplexed detection of Alzheimer’s disease Core biomarkers using PAapt-AuNPs conjugates. ACS Sensors 4(8):2140–2149.

    CAS  Article  Google Scholar 

  34. 34.

    Zhou J, Meng L, Ye W, Wang Q, Geng S, Sun C (2018) A sensitive detection assay based on signal amplification technology for Alzheimer's disease's early biomarker in exosome. Anal Chim Acta 1022(31):124–130.

    CAS  Article  Google Scholar 

  35. 35.

    Zhu X, Zhang N, Zhang Y, Liu B, Chang Z, Zhou Y, Xu M (2018) A sensitive gold nanoparticle-based aptasensor for colorimetric detection of Aβ 1–40 oligomers. Anal Methods 10(6):641–645.

    CAS  Article  Google Scholar 

Download references


This work was financially supported by 2018 Academic New Seedling Cultivation and Innovation Exploration Project of Guizhou Provincial Science and Technology Department (No. [2018]5784-05), and Ph. D. Foundation of Zunyi Normal College (No. BS[2019]10).

Author information



Corresponding author

Correspondence to Hong-Xia Ren.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material


(DOCX 330 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Ren, HX., Miao, YB. & Zhang, Y. An aptamer based fluorometric assay for amyloid-β oligomers using a metal-organic framework of type Ru@MIL-101(Al) and enzyme-assisted recycling. Microchim Acta 187, 114 (2020).

Download citation


  • Luminescent metal-organic framework
  • Enzyme assisted target recycling
  • Tracking agent
  • Homogeneous assay
  • Turn-on fluorometry