Microchimica Acta

, Volume 182, Issue 7–8, pp 1439–1446 | Cite as

Antibody-modified iron oxide nanoparticles for efficient magnetic isolation and flow cytometric determination of L. pneumophila

  • Maarten Bloemen
  • Carla Denis
  • Miet Peeters
  • Luc De Meester
  • Ann Gils
  • Nick Geukens
  • Thierry Verbiest
Original Paper
First Online:
Received:
Accepted:

Abstract

We report on the design of superparamagnetic nanoparticles capable of selectively isolating targeted bacteria (Legionella pneumophila, serogroup 1) from aqueous solutions. The surface of magnetite nanoparticles (NP) was functionalized with a heterobifunctional poly(ethylene glycol) ligand containing reactive groups for covalent coupling of polyclonal antibodies against L. pneumophila. These bioconjugates were used to label and magnetically isolate L. pneumophila. Flow cytometry revealed high separation and efficiency in this regard. The strain specificity and efficiency of the magnetic NP was tested with recombinant strains of E. coli (expressing the red fluorescent protein) and L. pneumophila (expressing the green fluorescent protein). The detection limit of the method (by flow cytometry) is 104 cells∙mL-1. The results indicate that the new multifunctional NPs are capable of selectively attracting pathogens from a complex mixture and with high efficiency. This, conceivably, paves the way to pre-concentration protocols for numerous other pathogens.

Graphical Abstract

Antibody-modified iron oxide nanoparticles were developed for selective magnetic isolation of L. pneumophila bacteria from water samples. A comprehensive flow cytometry study was performed for the quantification of the bacteria.

Keywords

Immunomagnetic isolation Iron oxide Nanoparticles Antibodies Superparamagnetism Heterobifunctional PEG Legionella pneumophila 
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Notes

Acknowledgments

We thank prof. Johan Billen for helping with the TEM measurements. This work was financially supported by grant G.0618.11 N of the Fund for scientific research Flanders (FWO-V) and the Agency for Innovation by Science and Technology in Flanders (IWT). M. Bloemen is grateful for support from the IWT.

Conflict of interest

The authors declare that they have no conflict of interest.

Compliance with ethical standards

All legal and ethical requirements for the welfare of animals were met by the Animalium Department of the KU Leuven.

Supplementary material

604_2015_1466_MOESM1_ESM.docx (1.8 mb)
ESM 1 (DOCX 1819 kb)

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Copyright information

© Springer-Verlag Wien 2015

Authors and Affiliations

  • Maarten Bloemen
    • 1
  • Carla Denis
    • 2
  • Miet Peeters
    • 3
  • Luc De Meester
    • 2
  • Ann Gils
    • 3
  • Nick Geukens
    • 4
  • Thierry Verbiest
    • 1
  1. 1.Department of ChemistryKU LeuvenHeverleeBelgium
  2. 2.Department of BiologyKU LeuvenLeuvenBelgium
  3. 3.Department of Pharmaceutical and Pharmacological SciencesKU LeuvenLeuvenBelgium
  4. 4.PharmAbsThe KU Leuven Antibody Center, KU LeuvenLeuvenBelgium

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