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. 2021 Jul 16;39(35):4988–5001. doi: 10.1016/j.vaccine.2021.07.034

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© 2021 The Authors

Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

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Fig. 1 — Constructs design and production of eVLPs expressing SARS-CoV-2 Spike protein. (a) Schematic representation of SARS-COV-2 S plasmid constructs. TM-CTD: Transmembrane cytoplasmic terminal domain. (b) Expression of SARS-CoV-2 S analyzed by Western-blot of SARS-CoV-2 eVLPs and recombinant proteins using a rabbit polyclonal Ab (pAb, upper panel) raised against SARS-CoV-2 RBD (Sinobiological) or COVID-19 convalescent human serum (HuCS, bottom panel). eVLPs produced with Gag plasmid only (Gag eVLPs) and recombinant SARS-CoV-2 S proteins were used as negative and positive controls respectively. (c) Detection of SARS-CoV-2 S by Western-blot (anti-RBD pAb) after overnight incubation at 37 °C with or without PNGase F (New England Biolabs) at 1000 and 2000 Units as indicated. MW: molecular weight ladder, r-S: recombinant native S, r-SP: recombinant prefusion S protein containing a mutated furin cleavage domain (RRAR → GSAS), replacement of 2 proline (KV → PP) and a trimerization domain.