Journal
ANALYTICA CHIMICA ACTA
Volume 1209, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.aca.2022.339863
Keywords
Upconversion nanoparticles; Antibody; Antibody-conjugated nanoparticles; Immunofluorescent analysis; Bioimaging
Categories
Funding
- Australian Research Council (ARC) through the Centre of Excellence for Nanoscale BioPhotonics [CE140100003]
- In-ternational Macquarie University Research Excellence Scholarship
- ARC Discovery Early Career Research Award [DE170100821]
- Macquarie University Research Fellowship
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A systematic approach was introduced to quantitatively evaluate the biological activity of antibody-conjugated UCNPs. An optimal antibody conjugation efficiency of approximately 70% was determined under specific conditions, providing a practical guideline for future studies on functionalized UCNPs.
Surface modification and functionalization is typically required to engineer upconversion nanoparticles (UCNPs) for biosensing and bioimaging applications. Nevertheless, despite various antibody conjugation methods having been applied to UCNPs, no consensus has been reached on the best choice, as the results from individual studies are largely unable to be compared due to inadequate assessment of the properties of the conjugated products. Here, we introduce a systematic approach to quantitatively evaluate the biological activity of antibodyconjugated UCNPs. We determine that the optimal antibody conjugation efficiency to our colominic acid polysaccharide-coated UCNPs via 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxy succinimide (EDC/NHS) coupling is approximately 70%, corresponding to 16 antibodies per nanoparticle of 63 nm hydrodynamic diameter, with on average 12 of the 16 antibodies maintaining their affinity to the target antigens. The binding ability of the antibody-conjugated UCNPs to the antigen was well preserved, as verified by enzymelinked immunosorbent assay (ELISA), flow cytometry, and cellular imaging. This is the first study to quantitate the active antibody binding capacity of polysaccharide coated UCNP nanoparticles, offering a practical guideline for benchmarking functionalised UCNPs in future studies.
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