Journal
MICROCHIMICA ACTA
Volume 186, Issue 6, Pages -Publisher
SPRINGER WIEN
DOI: 10.1007/s00604-019-3466-x
Keywords
Amplification; Bioconjugation; Nucleic acid analysis; Mini-emulsion polymerisation; Surface functionalisation; Magnetic separation; Biosensing; Luminescence; Lanthanide-doped
Categories
Funding
- Australian Research Council [LP140100462]
- Australian Research Council [LP140100462] Funding Source: Australian Research Council
Ask authors/readers for more resources
The authors report on a simplified approach to encapsulate upconversion nanoparticles (UCNPs) in polystyrene spheres by mini-emulsion polymerisation. The resulting particles (PS-UCNP) are hydrophilic, stable and suitable for biomolecular recognition and biosensing applications. Also, a strategy was developed for bioconjugation of antibodies onto the surface of the PS-UCNPs by using the bifunctional fusion protein linker-protein G (LPG). LPG mediates the functionalisation of PS-UCNPs with antibodies against digoxigenin allowing for specific labelling of convective PCR (cPCR) amplicons. Lambda DNA was amplified using cPCR on a heat block for 30min using the digoxigenin labelled forward and biotin labelled reverse primers. The antibody functionalised PS-UCNPs bind to the digoxigenin end of the cPCR amplicons. Finally, the streptavidin labelled magnetic beads were used to selectively capture the PS-UCNP-labelled cPCR amplicons and the upconversion signal was detected at 537nm under 980nm excitation. This sandwich approach enables direct recognition of the target lambda DNA with a detection limit of 10(3) copies L-1. The upconversion signal decreased proportionally to the concentration of the lambda DNA with a linear response between 10(7) and 10(3) copies of DNA.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available