Purification of quantum dot-based bioprobes via high-performance size exclusion chromatography

Talanta. 2016 Oct 1:159:64-73. doi: 10.1016/j.talanta.2016.05.078. Epub 2016 May 31.

Abstract

Due to excellent optical properties, quantum dots (QDs) have been widely applied to sensing, labeling, and imaging. For the fabrication of QD-based bioprobes, purification is usually the crucial step. Hydrophilic octylamine grafted polyacrylic acid modified QDs (OPA-QDs) were prepared, and purified by high-performance size exclusion chromatography (HPSEC) to remove excess OPA and aggregated QDs. The percentage of suspended agglomerates of OPA-QDs in the unpurified OPA-QDs increases from 4% to 31% through a year, but the purified OPA-QDs of the same batch possess excellent colloidal stability for at least one year. Subsequently, QD-based bioprobes were fabricated by the conjugation between QDs and streptavidin (SA) or antibody (IgG), generating QD-SA and QD-IgG, respectively, which were purified via HPSEC. Finally, the resulting QD-SA and QD-IgG were adopted to detect tumour markers on slices and showed specific positive signals without nonspecific adsorption, which was contrary to the unpurified QD-IgG. Thus, the HPSEC-coupled system proposed in the current work is potent and universal for the generation of purified and monodisperse QD-based bioprobes, which is promising in the nanobiodetection field.

Keywords: Bioprobes; High-performance size exclusion chromatography; Purification; Quantum dots.

MeSH terms

  • Antibodies / metabolism
  • Biomarkers, Tumor / metabolism
  • Biosensing Techniques*
  • Chemistry Techniques, Analytical / methods*
  • Chromatography, Gel*
  • Humans
  • Quantum Dots / chemistry*
  • Streptavidin / metabolism

Substances

  • Antibodies
  • Biomarkers, Tumor
  • Streptavidin