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. 2020 May 18;5:41. doi: 10.1038/s41541-020-0190-9

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© The Author(s) 2020

Open Access This 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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 1 — a Production of adjuvanted virosome-P1 (MYM-V111) and virosome-rgp41 (MYM-V112): Step 1, inactivated influenza A/H1N1 are solubilized with detergent; Step 2, nucleocapsides are discarded; Step 3, the viral membrane lipids with the native influenza hemagglutinin (HA) and neuraminidase (NA) are recovered; Steps 4a and 4b, synthetic lipids with 3M-052 adjuvant and antigen P1 or rgp41 are mixed with isolated viral membrane components and trehalose; Step 5, virosomes-P1 (pink rod) and virosome-rgp41 (blue rod) are gradually assembled in vitro during the detergent removal. Each virosome is then diluted and mixed together to generate the HIV-1 liquid vaccine MYM-V202. Universal T help provided by HA/NA. b Amnis® ImageStream on fluorescent Dil dye-labeled virosomes (in yellow) to visualize particles. The liquid virosome population contains mostly single particles (upper left image). Reconstituted powders contain also a major population of single particles, but a minor population of few small virosome clusters and bigger aggregates are also observed. Images were enlarged in Powerpoint because original AMNIS images are only tiny dots. c Mean particle size and population distribution monitored by NTA for the liquid bulk vaccine (MYM-V202, upper left panel), the reconstituted sublingual tablet (MYM-V212, upper right panel), the reconstituted nasal powder (MYM-V222, lower left panel), the reconstituted oral powder (MYM-V232, lower right panel). Black arrows identify the population of small clusters 200–300 nm and aggregates >300 nm, using arbitrary cut-off.