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. 2020 Apr 15;13:36. doi: 10.1186/s13045-020-00863-9

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© The Author(s) 2020

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 — T22-GFP-H6-Auristatin nanoconjugates characterization and stability. a T22-GFP-H6 nanoparticle conjugation with maleimide functionalized Monomethyl Auristatin E (MMAE). b MALDI-TOF mass spectrometry of wild-type T22-GFP-H6 nanoparticles and T22-GFP-H6-Auristatin nanoconjugates (top) and free MMAE before and after nanoconjugates re-purification (bottom). Each peak corresponds to the covalent addition (+ 911 Da) of a single MMAE molecule (top). Red boxes indicate free Auristatin mass spectrometry spectra (bottom). c Hydrodynamic volume size distribution of wild-type T22-GFP-H6 nanoparticles (red) and T22-GFP-H6-Auristatin nanoconjugates (blue) determined by dynamic light scattering. Samples were analyzed in triplicate and data represented as mean +/− SE. PDI indicates polydispersion index. d Average molar mass distribution of wild-type T22-GFP-H6 nanoparticles (red) and T22-GFP-H6-Auristatin nanoconjugates (blue) determined by size exclusion chromatography coupled to multi-angle light scattering (SEC-MALS). e Representative electron microscopy (FESEM) images of T22-GFP-H6-Auristatin nanoconjugates presented in two magnifications (see inset). Scale bars indicate 100 nm. In the bottom, the quantitative average size of nanoconstructs determined by image analysis and shown as mean ± SE. f Proteolytic stability of T22-GFP-H6-Auristatin nanoconjugates in human serum at different incubation times up to 24 h analyzed by western blot immunodetection with a monoclonal anti-His antibody. “S” indicates human serum control. MW, molecular weight; SE, standard error