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Label-Free Quantification of DNA Loading on Centrifugation-Resistant Spherical Nucleic Acids.

Seungheon Lee1, Subrata Pandit1, Gabriel Gilman1

  • 1Department of Chemistry, The University of Texas at Austin, Austin, Texas 78712, United States.

Analytical Chemistry
|June 2, 2025
PubMed
Summary
This summary is machine-generated.

Accurately quantifying DNA on spherical nucleic acids (SNAs) is vital for biosensing and therapy. A new, rapid, label-free method using DEAE-functionalized beads offers a faster, more accessible alternative for DNA loading determination.

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Area of Science:

  • Nanotechnology
  • Bioconjugation Chemistry
  • Analytical Chemistry

Background:

  • Accurate DNA loading on spherical nucleic acids (SNAs) is crucial for their performance in biosensing and therapeutic applications.
  • Existing methods for DNA quantification on SNAs are often time-consuming, labor-intensive, require sample loss, or are incompatible with centrifugation-resistant SNAs.
  • Many current techniques rely on expensive labels or hazardous reagents, limiting their accessibility and broad applicability.

Purpose of the Study:

  • To develop a simple, rapid, and label-free method for determining DNA loading on various types of SNAs.
  • To provide a versatile alternative to traditional DNA quantification techniques that are ineffective or inefficient for certain SNA constructs.
  • To establish a reliable method applicable to centrifugation-resistant SNAs with diverse core materials and DNA characteristics.

Main Methods:

  • Utilized diethylaminoethyl (DEAE)-functionalized beads to selectively bind free DNA based on ionic strength.
  • Developed a protocol for the selective elution of unbound DNA from SNA-bound DNA using adjustments in ionic strength.
  • Quantified eluted free DNA using standard laboratory equipment like UV-vis spectrophotometry or plate readers to calculate DNA loading per nanoparticle.

Main Results:

  • Successfully demonstrated the method's effectiveness on centrifugation-resistant SNAs with metallic or protein-based cores.
  • Validated the method across various DNA lengths, sequences, and densities, showing broad applicability.
  • Achieved accurate DNA loading determination in under 15 minutes, significantly faster than conventional methods.

Conclusions:

  • The DEAE-bead based method provides a fast, reliable, and label-free approach for quantifying DNA loading on SNAs.
  • This technique overcomes limitations of traditional methods, particularly for centrifugation-resistant SNAs.
  • The simplicity and broad applicability make it a valuable tool for nanoparticle-based research and development.