Journal: Human Vaccines & Immunotherapeutics
Article Title: Characterization and immunogenicity assessment of MERS-CoV pre-fusion spike trimeric oligomers as vaccine immunogen
doi: 10.1080/21645515.2024.2351664
Figure Lengend Snippet: Expi293F based mammalian expression system produced soluble trimeric prefusion MERS-CoV Spike. (a) schematic representation of color-coded expression cassette used in mammalian (pcDNA3.1 (1)) expression vector; blue box represents signal peptide sequence at N-terminal domain for mammalian expression; light-green box represents human-codon-optimized gene encoding M-CoV-S sequences from 16–1291 amino acids taken from England 1 strain; and at C-terminal domain dark-green box represents T4 fibritin trimerization domain and pink box for 8×His-tag. (b)12% reducing SDS-PAGE of purified M-CoV-S (Lane1: Marker, Lane 2: M-CoV-S). (c) Western blot image of inhouse M-CoV-S and commercial MERS-RBD (#10621-CV, R&D systems) probed with anti-M-CoV-S mouse polyclonal serum (Lane 1, 4: Marker, Lane 2: In-house M-CoV-S, Lane 3: recombinant MERS-RBD). (d) Blue native-PAGE image of purified M-CoV-S protein. Lane 1: native Marker, Lane 2: native M-CoV-S. (e) Purified M-CoV-S protein was subjected to de-glycosylation by using PNGase F enzyme (Lane 1: Marker, Lane 2: PNGase treated M-CoV-S, Lane 3: untreated M-CoV-S).
Article Snippet: Expi293F based mammalian expression system produced soluble trimeric prefusion MERS-CoV Spike. (a) schematic representation of color-coded expression cassette used in mammalian (pcDNA3.1 (1)) expression vector; blue box represents signal peptide sequence at N-terminal domain for mammalian expression; light-green box represents human-codon-optimized gene encoding M-CoV-S sequences from 16–1291 amino acids taken from England 1 strain; and at C-terminal domain dark-green box represents T4 fibritin trimerization domain and pink box for 8×His-tag. (b)12% reducing SDS-PAGE of purified M-CoV-S (Lane1: Marker, Lane 2: M-CoV-S). (c) Western blot image of inhouse M-CoV-S and commercial MERS-RBD (#10621-CV, R&D systems) probed with anti-M-CoV-S mouse polyclonal serum (Lane 1, 4: Marker, Lane 2: In-house M-CoV-S, Lane 3: recombinant MERS-RBD). (d) Blue native-PAGE image of purified M-CoV-S protein.
Techniques: Expressing, Produced, Plasmid Preparation, Sequencing, SDS Page, Purification, Marker, Western Blot, Recombinant, Blue Native PAGE