Evaluating binding avidities of populations of heterogeneous multivalent ligand-functionalized nanoparticles

ACS Nano. 2014 Jun 24;8(6):5600-9. doi: 10.1021/nn406455s. Epub 2014 May 13.

Abstract

Ligand-functionalized, multivalent nanoparticles have been extensively studied as targeted carriers in biomedical applications for drug delivery and imaging. The chemical synthesis method used, however, generates nanoparticles that are heterogeneous with respect to the number of ligands on each nanoparticle. This article examines the role this heterogeneity in ligand number plays in multivalent interactions between nanoparticle ligands and targeted receptors. We designed and synthesized a model heterogeneous multivalent nanoparticle system and developed a unique kinetic analysis to quantify the avidity interactions. This system used mono-dispersed poly(amidoamine) (PAMAM) dendrimers that were then chemically functionalized with ssDNA oligonucleotides as to yield the heterogeneous nanoparticle platform (ligand valencies n = 1.7, 3.1, 6), and employed complementary oligonucleotides as targeted receptors on a surface plasmon resonance (SPR) biosensor to evaluate the multivalent binding of the nanoparticle population. Kinetic analysis of both parallel initial rate and dual-Langmuir analyses of SPR binding curves was performed to assess avidity distributions. We found that batches of multivalent nanoparticles contain both fast- and slow-dissociation subpopulations, which can be characterized as having "weak" and "strong" surface interactions ("binding"), respectively. Furthermore, we found that the proportion of "strong" binders increased as a function of the mean oligonucleotide valence of the nanoparticle population. These analyses allowed an assessment of how avidity distributions are modulated by the number of functionalized ligands and suggested that there are threshold valences that differentiated fast- and slow-dissociation nanoparticles.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibody Affinity
  • Biosensing Techniques
  • Computer Simulation
  • DNA, Single-Stranded / chemistry
  • Dendrimers / chemistry
  • Kinetics
  • Ligands*
  • Macromolecular Substances
  • Nanoparticles / chemistry*
  • Nanotechnology / methods*
  • Oligonucleotides / chemistry
  • Polyamines / chemistry*
  • Protein Binding
  • Structure-Activity Relationship
  • Surface Plasmon Resonance

Substances

  • DNA, Single-Stranded
  • Dendrimers
  • Ligands
  • Macromolecular Substances
  • Oligonucleotides
  • Poly(amidoamine)
  • Polyamines