Detectio ion of f miRN iRNA by Activ tive Ca Capture + Dig igit - - PowerPoint PPT Presentation

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Detectio ion of f miRN iRNA by Activ tive Ca Capture + Dig igit - - PowerPoint PPT Presentation

Apr 09, 2024 24 likes •105 views

Detectio ion of f miRN iRNA by Activ tive Ca Capture + Dig igit ital l Co Countin ing (A (AC+ C+DC) ) Micr icrosc scopy Taylor Canady Carl R. Woese Fellow @ UIUC 1 Liq iquid id Bio iopsy: Bio ioflu luid id Bio iomarker r

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Detectio ion of f miRN iRNA by Activ tive Ca Capture + Dig igit ital l Co Countin ing (A (AC+ C+DC) ) Micr icrosc scopy

Taylor Canady Carl R. Woese Fellow @ UIUC

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Clin Cancer Res; 23(10) 2017

Op Opportunity of

  • f Liq

Liquid Bi Biopsy

  • Non-invasive
  • Early detection & quantification
  • Sort mutation types
  • Fast monitor and improve treatment

Cha Challe lenges of

  • f Liq

Liquid Bi Biopsy

  • Targets  Disease  Patient Diversity
  • Sequence similarity (SNPs)
  • Concentration & matrix
  • Build POC assay compatibility (time, cost, ...)

Des Desig ign Co Constr traints

  • Broad single base selectivity (DNA Nano)
  • Extreme sensitivity (PC Biosensors)
  • Wide dynamic range
  • No amp, no wash
  • Assay time: minutes to few hours

Liq iquid id Bio iopsy: Bio ioflu luid id Bio iomarker r Detectio ion

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Engin ineeri ring Tools ls for r li liquid id Bio iopsy of miRN iRNA Bio iomarkers Photonic ic Cry Crystal l Bio Biosensors DNA Nanotechnolo logy

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ΔI (PIV) Δλ (PWV)

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625 Wavelength (nm) Reflection (A.U.)

Peak Wavelength Value (PWV) Peak Intensity Value (PIV) Measure

λ θ

400 nm Evanescent Field |E|2(v2/m2) Computer simulation: Finite-Difference Time-Domain (FDTD)

Prin rincip iple le of f Photonic ic Cry Crystal l Nan anopartic icle le Se Sensin ing

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SLIDE 5

400 nm

25 µm

Digital Resolution of AuNP “Tags”: Detection Method

Hybrid causes drop in PC reflectance peak intensity & shift in wavelength Allows for digital resolution of particles

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Canady et al. PNAS, 2019

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Activ tive Ca Capture + Dig igit ital l Co Countin ing (A (AC+ C+DC) ) miRN iRNA Dia iagnostic ics

Canady et al. PNAS, 2019

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Background 0.1 fM 1 fM 10 fM 100 fM 1 pM miR-375 + AuNPs 120 mins 90 mins 60 mins 30 mins 1 min miR-375 + AuNPs 10 pM

625.3 625.0 25 µm

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2 hour endpoint Canady et al. PNAS, 2019

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SLIDE 8

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Highlights

Photonic Crystal Bios iosensin ing: Enhanced E-fie ield ld @ resonance  nan anoparticle detection Toe

  • ehold

ld DN DNA Probes: Optimize hyb ybrid idization by by ∆GRXN ≅ 0  si single le base ase se sele lectiv ivity

AC+DC Ass ssay: : Co Combin ine AuNP-DNA probe (s (selec lectiv ivit ity) ) & PC C bio iose sensors (s (sensit itiv ive) ) for r rapid id, no ampli lifi ficatio ion, and no wash miRN iRNA detectio ion AC+DC Data: miRN iRNA detectio ion with ith 106 dynamic ic range (1 (100 aM aM lo lowest) Sin Single le base se selec lectiv ivit ity across 22mer miRN iRNA (5 (5 example les) Detectio ion in in compli licated background (excess non-target RNA) Futu ture: Micr icroflu fluid idic ic im impla lantatio ion, addit itio ional l photonic ic-mic icroscopes