Molecular-receptor-specific, non-toxic, near-infrared-emitting Au cluster-protein nanoconjugates for targeted cancer imaging

Nanotechnology. 2010 Feb 5;21(5):055103. doi: 10.1088/0957-4484/21/5/055103. Epub 2009 Dec 21.

Abstract

Molecular-receptor-targeted imaging of folate receptor positive oral carcinoma cells using folic-acid-conjugated fluorescent Au(25) nanoclusters (Au NCs) is reported. Highly fluorescent Au(25) clusters were synthesized by controlled reduction of Au(+) ions, stabilized in bovine serum albumin (BSA), using a green-chemical reducing agent, ascorbic acid (vitamin-C). For targeted-imaging-based detection of cancer cells, the clusters were conjugated with folic acid (FA) through amide linkage with the BSA shell. The bioconjugated clusters show excellent stability over a wide range of pH from 4 to 14 and fluorescence efficiency of approximately 5.7% at pH 7.4 in phosphate buffer saline (PBS), indicating effective protection of nanoclusters by serum albumin during the bioconjugation reaction and cell-cluster interaction. The nanoclusters were characterized for their physico-chemical properties, toxicity and cancer targeting efficacy in vitro. X-ray photoelectron spectroscopy (XPS) suggests binding energies correlating to metal Au 4f(7/2) approximately 83.97 eV and Au 4f(5/2) approximately 87.768 eV. Transmission electron microscopy and atomic force microscopy revealed the formation of individual nanoclusters of size approximately 1 nm and protein cluster aggregates of size approximately 8 nm. Photoluminescence studies show bright fluorescence with peak maximum at approximately 674 nm with the spectral profile covering the near-infrared (NIR) region, making it possible to image clusters at the 700-800 nm emission window where the tissue absorption of light is minimum. The cell viability and reactive oxygen toxicity studies indicate the non-toxic nature of the Au clusters up to relatively higher concentrations of 500 microg ml(-1). Receptor-targeted cancer detection using Au clusters is demonstrated on FR(+ve) oral squamous cell carcinoma (KB) and breast adenocarcinoma cell MCF-7, where the FA-conjugated Au(25) clusters were found internalized in significantly higher concentrations compared to the negative control cell lines. This study demonstrates the potential of using non-toxic fluorescent Au nanoclusters for the targeted imaging of cancer.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Carrier Proteins / metabolism*
  • Cell Line
  • Cell Line, Tumor
  • Cell Survival
  • Flow Cytometry
  • Folate Receptors, GPI-Anchored
  • Folic Acid / chemistry*
  • Folic Acid / metabolism
  • Formazans
  • Gold / chemistry*
  • Gold / metabolism
  • Humans
  • Luminescent Measurements
  • Microscopy, Atomic Force
  • Microscopy, Electron, Transmission
  • Microscopy, Fluorescence
  • Mouth Neoplasms / diagnosis*
  • Mouth Neoplasms / metabolism
  • Mouth Neoplasms / pathology
  • Nanocomposites / chemistry*
  • Photoelectron Spectroscopy
  • Reactive Oxygen Species / metabolism
  • Receptors, Cell Surface / metabolism*
  • Serum Albumin, Bovine / chemistry
  • Spectroscopy, Fourier Transform Infrared
  • Tetrazolium Salts

Substances

  • Carrier Proteins
  • Folate Receptors, GPI-Anchored
  • Formazans
  • Reactive Oxygen Species
  • Receptors, Cell Surface
  • Tetrazolium Salts
  • MTT formazan
  • Serum Albumin, Bovine
  • Gold
  • Folic Acid