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US-20260126436-A1 - AMPLIFYING BIOMOLECULAR SENSING THROUGH ACOUSTICALLY ENHANCED REACTIONS

US20260126436A1US 20260126436 A1US20260126436 A1US 20260126436A1US-20260126436-A1

Abstract

The disclosed subject matter provides methods and apparatus for detecting a target analyte in a sample. The method includes providing nanoparticles functionalized with one or more recognition molecules configured to bind specifically to the target analyte; exposing the sample containing the target analyte and the functionalized nanoparticles to an acoustic field comprising a surface standing acoustic wave (SSAW); inducing aggregation of the nanoparticles mediated by the target analyte under acoustic stimulation of SSAW; and detecting the aggregation of the nanoparticles as an indication of the presence of the target analyte.

Inventors

  • Oleg Gang
  • Feiyue Teng
  • Zohar Albert ARNON

Assignees

  • THE TRUSTEES OF COLUMBIA UNIVERSITY IN THE CITY OF NEW YORK
  • BROOKHAVEN SCIENCE ASSOCIATES, LLC

Dates

Publication Date
20260507
Application Date
20251105

Claims (20)

  1. 1 . A method of detecting a target analyte in a sample, comprising: providing nanoparticles, functionalized with one or more recognition molecules configured to bind specifically to the target analyte, to the sample; exposing the sample with the target analyte and the functionalized nanoparticles to an acoustic field comprising a surface standing acoustic wave (SSAW); inducing aggregation of the nanoparticles mediated by the target analyte under acoustic stimulation of the SSAW; and detecting the aggregation of the nanoparticles as an indication of presence of the target analyte.
  2. 2 . The method of claim 1 , wherein the acoustic field is generated by applying an electrical signal to a piezoelectric substrate.
  3. 3 . The method of claim 1 , wherein the acoustic field is configured to generate acoustic excitation having a frequency between about 1 MHz and about 50 MHz.
  4. 4 . The method of claim 1 , wherein the acoustic stimulation is applied in pulses having a duration of about 10 ms to about 100 ms with a period of about 0.1 s to about 5 s.
  5. 5 . The method of claim 1 , wherein the recognition molecules comprise at least one of antibodies, antibody fragments, aptamers, nucleic acids, or enzymes.
  6. 6 . The method of claim 1 , wherein the target analyte comprises at least one of DNA linker, protein, nucleic acid, pathogen, toxin, metabolite, or biomarker associated with a disease state.
  7. 7 . The method of claim 1 , wherein the detecting comprises at least one of optical detection, dynamic light scattering, dark-field microscopy, spectrophotometry, or electron microscopy.
  8. 8 . The method of claim 1 , wherein the SSAW is generated by a plurality of interdigital transducers (IDTs) patterned on a piezoelectric substrate.
  9. 9 . The method of claim 1 , further comprising tuning pulse duration and acoustic period to adjust aggregation efficiency.
  10. 10 . The method of claim 1 , wherein the nanoparticles are functionalized with two or more recognition molecules binding to non-overlapping epitopes of the analyte.
  11. 11 . The method of claim 1 , wherein the detecting comprises monitoring a plasmonic shift in optical spectra associated with aggregation of the nanoparticles.
  12. 12 . An apparatus for detecting a target analyte in a sample, comprising: a reaction chamber having a capillary configured to receive a sample containing a target analyte and nanoparticles functionalized with recognition molecules; a piezoelectric substrate operatively coupled to the reaction chamber; an electrical signal generator configured to apply an electrical signal to the piezoelectric substrate to generate a surface standing acoustic wave (SSAW) within the capillary; and a detector configured to measure an aggregation of the nanoparticles mediated by the target analyte.
  13. 13 . The apparatus of claim 12 , wherein the detector comprises at least one of an optical detector, spectrophotometer, dark-field microscope, or dynamic light scattering instrument.
  14. 14 . The apparatus of claim 12 , wherein the nanoparticles comprise gold nanoparticles conjugated to two or more antibodies recognizing specific epitopes of the target analyte.
  15. 15 . The apparatus of claim 12 , wherein the piezoelectric substrate comprises lithium niobate (LiNbO 3 ).
  16. 16 . The apparatus of claim 12 , wherein the capillary comprises a glass capillary mounted on the piezoelectric substrate.
  17. 17 . The apparatus of claim 12 , wherein the detector is configured to detect changes in hydrodynamic radius of nanoparticle clusters using dynamic light scattering.
  18. 18 . The apparatus of claim 12 , wherein the detector is configured to output a visual indication of aggregation at analyte concentrations at least 100-fold lower than detection in absence of acoustic stimulation.
  19. 19 . The apparatus of claim 12 , wherein the electrical signal generator is configured to apply electrical signals to an interdigital transducer to generate counter-propagating acoustic waves.
  20. 20 . The apparatus of claim 19 , wherein the electrical signals are configured to vary at least one of frequency, amplitude, or duty cycle to adaptively tune the SSAW.

Description

CROSS-REFERENCE TO RELATED APPLICATION This Application claims priority to U.S. Provisional Patent Application Ser. No. 63/716,794, filed Nov. 6, 2024, which is hereby incorporated by reference in its entirety. GRANT INFORMATION This invention was made with government support under DE-SC0012704 awarded by the U.S. Department of Energy and W911NF-22-2-0111 awarded by the Army Research Office. The government has certain rights in the invention. BACKGROUND The disclosed subject matter relates generally to biosensing and diagnostic technologies, and more particularly to techniques for amplifying biomolecular sensing through acoustically enhanced reactions. Point-of-care diagnostic techniques are increasingly important tools for rapid detection of molecular biomarkers in healthcare, food safety, environmental monitoring, and related fields. A limitation of certain methods is their sensitivity, which can be constrained by the inherent kinetics of molecular recognition processes. For example, molecular recognition between a target analyte and a receptor (e.g., antibody, aptamer, or enzyme) often proceeds slowly and can require relatively high analyte concentrations to achieve reliable detection due to a kinetic barrier, thereby limiting applicability in low-abundance biomarker scenarios, such as early-stage disease diagnostics. Therefore, there is a need for developing techniques to overcome the limitations in biomolecular recognition in biosensors. SUMMARY The disclosed subject matter provides techniques for a broad and reliable detection of target biomolecules, enabling the detection of target biomolecules of small cluster sizes and low concentrations. In particular, the disclosed subject matter employs acoustic wave energy to accelerate molecular recognition kinetics in nanoparticle-based biosensors, thereby significantly increasing detection sensitivity across a wide range of target analytes and sensing platforms. An example method comprises providing nanoparticles functionalized with one or more recognition molecules configured to bind specifically to the target analyte, exposing the sample containing the target analyte and the functionalized nanoparticles to an acoustic field comprising a surface standing acoustic wave (SSAW), inducing aggregation of the nanoparticles mediated by the target analyte under acoustic stimulation of the SSAW, and detecting the aggregation of the nanoparticles as an indication of the presence of the target analyte. In certain embodiments, the acoustic field is generated by applying an electrical signal to a piezoelectric substrate, such as lithium niobate (LiNbO3), through a plurality of interdigital transducers (IDTs). The acoustic field can be configured to generate excitation at frequencies between about 1 MHz and about 50 MHz. In certain embodiments, acoustic stimulation is applied in pulses having a duration of about 10 ms to about 100 ms with a period of about 0.1 s to about 5 s. The pulse duration and acoustic period can be tuned to adjust aggregation efficiency. In certain embodiments, the recognition molecules conjugated to the nanoparticles can include at least one of antibodies, antibody fragments, aptamers, nucleic acids, or enzymes. The target analyte can include at least one of DNA linker, protein, nucleic acid, pathogen, toxin, metabolite, or biomarker associated with a disease state. In certain embodiments, nanoparticles are functionalized with two or more recognition molecules that bind to non-overlapping epitopes of the analyte. In certain embodiments, the detecting comprises at least one of optical detection, dynamic light scattering, dark-field microscopy, spectrophotometry, or electron microscopy. The SSAW is generated by the IDTs patterned on a piezoelectric substrate. The method further can comprise tuning pulse duration and acoustic period to adjust aggregation efficiency. The detecting comprises monitoring a plasmonic shift in optical spectra associated with aggregation of the nanoparticles. In another aspect, the disclosed subject matter provides an apparatus for detecting a target analyte in a sample. An example apparatus can include a reaction chamber comprising a capillary configured to receive a sample containing a target analyte and nanoparticles functionalized with recognition molecules; a piezoelectric substrate operatively coupled to the reaction chamber; an electrical signal generator configured to apply an electrical signal to the piezoelectric substrate to generate a SSAW within the capillary; and a detector configured to measure aggregation of the nanoparticles mediated by the target analyte. In certain embodiments, the detector can include an optical detector, spectrophotometer, dark-field microscope, or dynamic light scattering instrument. The detector can be configured to detect changes in hydrodynamic radius of nanoparticle clusters or to output a visual indication of aggregation. The nanoparticles can include gold nanoparticles conjugated