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A simple and sensitive biosensor for rapid detection of nanoparticles in water

  • Prasanna Bhomkar
  • Greg Goss
  • David S. Wishart
Research Paper

Abstract

Advances in nanoscience have led to a greater use of engineered nanoparticles (ENPs) in numerous applications. Due to their small size and unique surface properties, ENPs have many desirable features. However, they also interact with living cells in potentially undesirable manners highlighting the need to develop improved detection systems to manage risks associated with their accidental occupational exposure or environmental release. However, the routine detection of ENPs has not yet been demonstrated, especially for aquatic environments. Using standard protein engineering techniques, we generated a protein-based biosensor that can sensitively detect negatively charged ENPs in aquatic matrices. In particular, we genetically engineered a green fluorescent protein with a poly-lysine tag (His-GFP-LYS) to facilitate its electrostatic interaction with commercially available negatively charged NPs. These 5–6-nm-sized NPs have metallic cores comprising gold, iron oxide, cerium oxide, and zinc oxide and are stabilized via poly-acrylic acid (PAA) coating. The interaction between the recombinant positively charged GFP and the PAA coating of the negatively charged NPs resulted in visually observable turbidity changes that were quantified using a portable spectrophotometer (NANODROP). These interactions were confirmed using dynamic light scattering and visualized using agarose native gel electrophoresis. This simple and portable system could detect ENPs resuspended in pure aqueous buffer (0.08 mg/L) and those resuspended in environmental matrices, such as pond water (0.6 mg/L). This detection system also sensed ENPs in the presence of moderate concentrations of natural organic matter that is ubiquitously present in surface waters. These results suggest that this biosensor system could be used for the routine, portable, and affordable detection of negatively-charged ENPs under environmentally relevant aquatic conditions.

Keywords

Engineered nanoparticles Biosensor Detection GFP Nanomaterial 

Notes

Acknowledgments

The authors wish to thank the NRC-NSERC-BDC Nanotechnology Initiative (National Research Council-Natural Sciences and Engineering Research Council-Business Development Bank of Canada) for financial support; and Vive Crop Protection for providing the nanomaterials.

Supplementary material

11051_2014_2253_MOESM1_ESM.docx (15 kb)
Supplementary material 1 (DOCX 14 kb)
11051_2014_2253_MOESM2_ESM.tif (147 kb)
Supplementary Fig. 1 Schematic illustration of the proposed interaction between positively charged His-GFP-LYS and negatively charged nanoparticles. Positively charged GFP is depicted as green cylinders with the “+” sign denoting the poly-lysine tail. Red circles with the “−” sign denote negatively charged nanoparticles. Panel a is a scenario wherein there is less protein available per nanoparticle compared to the scenario in panel b. Due to an excess of protein in the latter scenario, some of the proteins can bridge adjacent nanoparticles via electrostatic interactions, eventually resulting in the formation of large agglomerates (TIFF 146 kb)
11051_2014_2253_MOESM3_ESM.tif (249 kb)
Supplementary Fig. 2 Analysis of filtration-assisted turbidimetric detection of His-GFP-LYS:Vive Nano gold interaction using native agarose gel electrophoresis. 500 μL of Vive Nano gold (resuspended in 100 mM sodium phosphate buffer pH 7) were centrifuged using 3 kDa MWCO filters, following which 2 μL of the concentrated fraction was mixed with 100 μL of His-GFP-LYS (0.7 mg/mL). The reaction mixture was incubated for 30 min following which twenty microliters of the reaction mix was loaded onto a 1 % Tris borate EDTA agarose gel. The different lanes refer to 1 His-GFP alone, 2 His-GFP + Vive nano gold (10 mg/L), 3 His-GFP-LYS alone. Lanes 48 refer to His-GFP-LYS mixed with Vive Nano gold (final concentrations: 20, 10, 5, 2.5, and 1.25 mg/L,). Lanes 913 refer to concentrated fractions of Vive Nano gold (final concentrations 1.25, 0.6, 0.3, 0.15, and 0.08 mg/L) (TIFF 249 kb)

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Copyright information

© Her Majesty the Queen in Rights of Canada  2014

Authors and Affiliations

  • Prasanna Bhomkar
    • 1
  • Greg Goss
    • 1
    • 3
    • 4
  • David S. Wishart
    • 1
    • 2
    • 3
  1. 1.National Institute for NanotechnologyEdmontonCanada
  2. 2.Department of Computing ScienceUniversity of AlbertaEdmontonCanada
  3. 3.Department of Biological SciencesUniversity of AlbertaEdmontonCanada
  4. 4.Office of Environmental NanosafetyUniversity of AlbertaEdmontonCanada

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