Direct conjugation of silicon nanoparticles with M13pVIII-engineered proteins to bacteria identification
- 61 Downloads
Pseudomonas aeruginosa and Staphylococcus aureus are two opportunistic human pathogens among the most common agents of nosocomial infections. The early detection plays an important role in health care, and pharmacological and biomedical sectors. The use of functional nanoparticles is promising candidate to create new materials and devices to improve diagnosis, prevention, and treatment of diseases in different fields of applications. In this work, we used phage-specific pVIII proteins, isolated from P9b and St.au9IVS5 phage clones, displaying exogenous peptide (QRKLAAKLT and RVRSAPSSS) to detect P. aeruginosa and S. aureus, respectively. These selective bioprobes were used in “one-step” functionalization of silicon nanoparticles (SiNPs) by pulsed laser ablation of silicon in an aqueous solution, containing phage-specific pVIII protein. The optical properties of the bioconjugates (pVIII–SiNPs) are examined by photoluminescence and UV–Vis spectroscopy. Furthermore, size distribution and ability of bioconjugates to bind its bacterial target has been investigated by scanning electron microscopy, scanning transmission electron microscopy, and epi-fluorescence microscope. Our results show that the bioconjugates are able to bind P. aeruginosa and S. aureus, respectively, within 30 min. Furthermore, the yellow–green photo-emissive properties, detected by epi-fluorescence microscopy, demonstrate their potential use as fluorescent probes silicon-based for in vitro applications.
- P9b pVIII–SiNPs
Bioconjugate of SiNPs (silicon nanoparticles) with pVIII-engineered protein isolated from P9b phage clone.
- St.au9IVS5 pVIII–SiNPs
Bioconjugate of SiNPs (silicon nanoparticles) with pVIII-engineered protein isolated from St.au9IVS5 phage clone.
The authors thank Dr. F. Barreca and Prof. F. Neri for help measurements with scanning electron microscopy operating in transmission mode (STEM).
LP, MGR, and DF performed isolation of specific pVIII protein from P9b and St.au9IVS5 phage clones; one-step synthesis of pVIII–SiNPs bioconjugates; binding of P. aeruginosa and S. aureus to pVIII–SiNPs complexes. S.S. and A.M.M. analyzed the pVIII–SiNPs bioconjugates samples after and post-binding of P. aeruginosa. LP, MGR, DF, SS, AMM, and SG discussed and analyzed data, and wrote the manuscript. All authors read and approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors report no conflicts of interest in this work.
- 2.K. Huang, H. Ma, J. Liu, S. Huo, A. Kumar, T. Wei, X. Zhang, S. Jin, Y. Gan, P.C. Wang, S. He, X. Zhang, X.-J. Liang, Size-dependent localization and penetration of ultrasmall gold nanoparticles in cancer cells, multicellular spheroids, and tumors in vivo. ACS Nano 6, 4483 (2012)CrossRefGoogle Scholar
- 9.P. Liu, L. Han, F. Wang, V.A. Petrenko, A. Liu, Gold nanoprobe functionalized with specific fusion protein selection from phage display and its application in rapid, selective and sensitive colorimetric biosensing of Staphylococcus aureus. Biosensors Bioelectron. 82, 195–203 (2016). https://doi.org/10.1016/j.bios.2016.03.075 CrossRefGoogle Scholar
- 10.M. Karlsson, U. Carlsson, Protein adsorption orientation in the light of fluorescent probes: mapping of the interaction between site-directly labeled human carbonic anhydrase II and silica nanoparticles. Biophys. J. 88, 3536–3544 (2005). https://doi.org/10.1529/biophysj.104.054809 ADSCrossRefGoogle Scholar
- 11.K. Bagga, A. Barchanski, R. Intartaglia, S. Dante, R. Marotta, A. Diaspro, C.L. Sajti, F. Brandi, Laser-assisted synthesis of Staphylococcus aureus protein-capped silicon quantum dots as bio-functional nanoprobes. Laser Phys. Lett. 10, 065603 (2013). https://doi.org/10.1088/1612-2011/10/6/065603 ADSCrossRefGoogle Scholar
- 12.G.T. Hermanson, Bioconjugate Techniques, 2nd edn. (Elsevier, Amsterdam, 2008). ISBN: 978-0-12-370501-3Google Scholar
- 20.C. Pfeiffer, C. Rehbock, D. Hu¨hn, C. Carrillo-Carrion, D. Jimenez de Aberasturi, V. Merk, S. Barcikowski, W.J. Parak, Interaction of colloidal nanoparticles with their local environment: the (ionic) nanoenvironment around nanoparticles is different from bulk and determines the physico-chemical properties of the nanoparticles. J. R. Soc. Interface 11, 20130931 (2014)CrossRefGoogle Scholar
- 24.C.F. Barbas, D.R. Burton, J.K. Scott, G.J. Silverman, Phage Display, A Laboratory Manual (Cold Spring Harbor Lab. Press, Woodbury, 2001)Google Scholar
- 30.P.K. Jayanna, D. Bedi, J.W. Gillespie, P. DeInnocentes, T. Wang, V.P. Torchilin, R.C. Bird, V.A. Petrenko, Landscape phage fusion protein-mediated targeting of nanomedicines enhances their prostate tumor cell association and cytotoxic efficiency. Nanomed. Nanotechnol. Biol. Med. 6, 538–546 (2010). https://doi.org/10.1016/j.nano.2010.01.005 CrossRefGoogle Scholar
- 37.C. Rehbock, V. Merk, L. Gamrad, R. Streubel, S. Barcikowski, Size control of laser-fabricated surfactant-free gold nanoparticles with highly diluted electrolytes and their subsequent bioconjugation. Phys. Chem. Chem. Phys. 15, 3057–3067 (2013). https://doi.org/10.1039/c2cp42641b CrossRefGoogle Scholar
- 38.A.A. Shemetov, I. Nabiev, A. Sukhanova, Molecular interaction of proteins and peptides with nanoparticles. ACNS Nano 6, 4585–4602 (2012)Google Scholar
- 42.F.X. Schmid, in Encyclopedia Life Sciences, Introductory Articles, R. Bridgewater (ed.) (Wiley, 2001), pp. 1–4. https://doi.org/10.1038/npg.els.0003142
- 47.S. Hamad, G.K. Podagatlapalli, V.S. Vendamani, S.V.S. Nageswara Rao, A.P. Pathak, S.P. Tewari, S. Venugopal Rao, Femtosecond ablation of silicon in acetone: tunable photoluminescence from generated nanoparticles and fabrication of surface nanostructures. J. Phys. Chem. C 118, 7139–7151 (2014). https://doi.org/10.1021/jp501152x CrossRefGoogle Scholar