Mymetics Corp (MYMX)

[Work Harder]

In addition, the S protein was covalently coupled via its His-tag to
a click chemistry lipid present in the virosomal membrane.

2.4. Virosomes Preparation

To the supernatant, the lipid 1,2-dioleoyl-sn-glycero-3-phosphocholine
(DOPC) (Merck) and the click chemistry lipid dicyclobenzooctyl phosphatidylethanolamine (DBCO-PE) (Avanti Polar Lipids, Alabaster, AL, USA) were dissolved in OEG, and added OEG was then removed by batch chromatography on polystyrene beads (BioBeads SM2) as described; resulting homogenous virosomes were sterilized by 0.22 µm filtration.

2-azidoethyl thiophosphodichlorate (ATPD) was synthesized and purified as described
(Jia, S. et al., 2020) by Acme Bioscience (Palo Alto, CA, USA). S protein was dialyzed against 50 mM HEPES pH 8.5 for 4 h and then mixed with ATPD at a 200:1 ratio of ATPD to protein for 1 h at RT. The product was dialyzed overnight against 2000 volumes of buffer (145 mM NaCl, 5 mM HEPES, 1 mM EDTA, pH 7.4). The resulting S-azide conjugate product was incubated with virosomes for at least 24 h at 25 ?C resulting in covalent coupling of S to virosomes through azide-DBCO-PE click chemistry.

3.3. Conjugation of SARS-CoV-2 Spike Protein to Virosomes

Purified S protein was covalently coupled to virosomes through DBCO-azide click
chemistry, and the presence of S protein on the virosomes through the exposure of key
epitopes on the protein and the binding of an ACE2-Fc were confirmed by ELISA (Figure 4). Results indicate that the S protein as displayed on the surface of the virosomes is capable of binding to the ACE2 receptor and is also recognized by CR3022 and by all the tested neutralizing antibodies toward various epitope clusters. This binding capacity is also preserved on virosomes-S stored at 4 ?C for 1 month, as similar IC50 values were obtained for t = 0 and t = 1 month, thus demonstrating an enhanced stability of virosomes-S.

These results suggest that S protein covalently coupled via its
His tag to a click chemistry lipid present in the virosomal membrane results in an oriented display of the protein and properly exposes its receptor binding domain (RBD) involved in ACE2 binding on target cells, thus, theoretically favoring the induction of relevant neutralizing antibodies toward RBD for blocking cell infection

Insect Cells for High-Yield Production of SARS-CoV-2 Spike Protein: Building a Virosome-Based COVID-19 Vaccine Candidate

https://pubmed.ncbi.nlm.nih.gov/35456687/

A SARS-CoV-2 Wuhan spike virosome vaccine induces superior neutralization breadth compared to one using the Beta spike

4 times

https://www.nature.com/articles/s41598-022-07590-w

From the Phospene post

Study investigates the production of a virosome-based COVID-19 vaccine candidate

The study

In a recent study published in Pharmaceutics, different signal peptides, baculovirus transfer vectors, cell lines, infection techniques, and formulation buffers were investigated with the purpose of building a scalable bioprocess to generate high-quality S protein for incorporation in a virosome-based COVID-19 vaccine candidate.

The stability, oligomeric state, and binding capability of the generated protein to the angiotensin-converting enzyme 2 (ACE2) receptor and selected neutralizing SARS-CoV-2 antibodies were all evaluated in depth. The S protein was also covalently linked to a click chemistry lipid in the virosomal membrane through its polyhistidine- (His)-tag.

https://www.news-medical.net/news/20220418/Study-investigates-the-production-of-a-virosome-based-COVID-19-vaccine-candidate.aspx

Recent trends in click chemistry as a promising technology for virus-related research

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7173221/

About the time Baylor showed up. Speaking of Baylor, it's almost November 2 years ago & Ronnie didn't pr what he should have & pr ed what he shouldn't have ?

PCI Biotech is in Norway right

Bavarian Nordic A/S (OMX: BAVA, OTC: BVNRY) Kvistgaard, Denmark

https://www.pcibiotech.no/news/pci-biotech-initiates-research-collaboration-with-bavarian-nordic