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. 2021 May 5;19:128. doi: 10.1186/s12951-021-00871-y

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© The Author(s) 2021

Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 1 — Characterization of tri- and bifunctional nanodrugs by UV–Vis spectrophotometry and binding assays to αvβ3, sTNF-R2 and anti-IL12 antibodies. a UV–Vis absorption spectra of nanodrugs. The dotted line and solid line correspond to uncoated or coated 25-nm gold nanoparticles, respectively. The TNF:IL12 molar ratio used to prepare the trifunctional nanodrugs is indicated in parentheses. b Binding of different nanodrugs (iso1Au, Au/TNF, Au/IL12, iso1Au/IL12, iso1Au/TNF, and iso1Au/TNF + IL12) to αvβ3, soluble TNF receptor Type-II (sTNF-R2, Enbrel), and anti-IL12 antibody (mAb C15.6) adsorbed onto nitrocellulose filters, as detected by silver staining of gold nanoparticles. The iso1Au/TNF + IL12 used in this experiment was prepared using a TNF:IL12 molar ratio of 4:1. c Binding of nanodrugs to microtiter plates coated with or without αvβ3, as detected using a biotinylated antibody anti-IL12 (mAb C17.8) or with anti-TNF antibodies followed by HRP-labeled streptavidin or HRP-labeled secondary antibodies, respectively. Schematic representation of the assays (upper panels). The results of a representative experiment are shown (mean ± SE of duplicates). The TNF:IL12 molar ratio used to prepare the nanodrugs is indicated in parentheses