Development of anti-TNFRSF-conjugated nanoparticle and in vitro evaluation of its immunostimulatory effect.

Immunotherapy is considered to be one of the most promising cancer treatments. However, applications of some promising immunotherapeutic, such as anti-fibroblast growth factor-inducible 14 (anti-Fn14), are limited due to their low bioactivity. Anti-Fn14 antibodies mimic TWEAK, a member of the tumor necrosis factor superfamily TNFSF ligands, which stimulate Fn14 (TNFRSF) which results in the induction of necrotic and apoptotic cell death.

It has previously shown that the bioactivity of such antibodies, could be improved by their crosslinking, that is immobilization of multiple antibodies in close proximity. In this work, we have developed a method to increase the activity of the anti-Fn14 antibodies by physical cross-linking on gold nanoparticles. We have developed and optimized the method for the preparation of gold nanoparticles, their functionalization with poly-ethylene glycol (PEG) linkers, and grafting of the antibodies on the surface.

We showed that antibodies can be successfully attached to nanoparticles without affecting their activity. Most importantly, the bioactivity measured of the grafted antibodies was increased in comparison to non-grafted antibodies as verified by the triggering HT-1080 cell line to produce the inflammatory cytokine IL-8 which was characterized by ELISA assay.

Our results suggest that conjugation of monoclonal IgG1 antibody on the inorganic nanoparticles is a very promising technique to boost the efficacy of the immunotherapeutic.

FUNDING:

This project has received funding from the European Union´s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 813871.

REFERENCES:

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