Cellular uptake of fluorophore-labeled glyco-DNA–gold nanoparticles

  • Katrin G. Witten
  • Julie Ruff
  • Anne Mohr
  • Dieter Görtz
  • Tobias Recker
  • Natalie Rinis
  • Claudia Rech
  • Lothar Elling
  • Gerhard Müller-Newen
  • Ulrich Simon
Research Paper

Abstract

DNA-functionalized gold nanoparticles (AuNP–DNA) were hybridized with complementary di-N-acetyllactosamine-(di-LacNAc, [3Gal(β1-4)GlcNAc(β1-]2)-modified oligonucleotides to form glycol-functionalized particles, AuNP–DNA–di-LacNAc. While AuNP–DNA are known to be taken up by cells via scavenger receptors, glycol-functionalized particles have shown to be taken up via asialoglycoprotein receptors (ASGP-R). In this work, the interaction of these new particles with HepG2 cells was analyzed, which express scavenger receptors class B type I (SR-BI) and ASGP-R. To study the contribution of these receptors as potential mediators for cellular uptake, receptor-blocking experiments were performed with d-lactose, a ligand for ASGP-R, Fucoidan, a putative ligand for SR-BI, and a SR-BI blocking antibody. Labeling with Cy5-modified DNA ligands enabled us to monitor the particle uptake by confocal fluorescence microscopy and flow cytometry, in order to discriminate the two putative pathways by competitive binding studies. While SR-BI-antibody and d-lactose had no inhibiting effects on particle uptake Fucoidan led to a complete inhibition. Thus, a receptor-mediated uptake by the two receptors studied could not be proven and therefore other uptake mechanisms have to be considered.

Keywords

Gold nanoparticles DNA Carbohydrates Cellular uptake Hepatocytes 

Supplementary material

11051_2013_1992_MOESM1_ESM.docx (652 kb)
Supplementary material 1 (DOCX 652 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Katrin G. Witten
    • 1
  • Julie Ruff
    • 1
  • Anne Mohr
    • 2
  • Dieter Görtz
    • 2
  • Tobias Recker
    • 2
  • Natalie Rinis
    • 2
  • Claudia Rech
    • 3
  • Lothar Elling
    • 3
  • Gerhard Müller-Newen
    • 2
  • Ulrich Simon
    • 1
  1. 1.Institute of Inorganic Chemistry and JARA - Fundamentals of Future Information TechnologyRWTH Aachen UniversityAachenGermany
  2. 2.Institute of Biochemistry and Molecular Biology, University Hospital AachenRWTH Aachen University52074 AachenGermany
  3. 3.Laboratory for Biomaterials, Institute of Biotechnology and Helmholtz-Institute for Biomedical EngineeringRWTH Aachen UniversityAachenGermany

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