Journal: NPJ Vaccines
Article Title: DS2 designer pre-fusion F vaccine induces strong and protective antibody response against RSV infection
doi: 10.1038/s41541-024-01059-9
Figure Lengend Snippet: A The pre-fusion and post-fusion conformational states of the RSV F glycoprotein are depicted, with colored antigenic sites (Ø, V, IV, III, II, and I) recognized by distinct groups of monoclonal antibodies (mAbs). B Specific mutations introduced to maintain the prefusion conformation in the three designer pre-F glycoproteins DS2, DS-Cav1, and SC-TM. C A schematic diagram illustrates the recombinant AdC68 vector expressing the three designer pre-F glycoproteins of RSV. Each glycoprotein was presented with signal peptide (SP), transmembrane (TM), and cytoplasmic tail (CT) from RSV A2 strain. The coding sequences of each designer pre-F were inserted into the E1 region of the AdC68 vector, controlled of the cytomegalovirus (CMV) promoter, and terminated by a bovine growth hormone (BGH) polyadenylation signal sequence. D A comparison of the expression and epitope display of various designer pre-F on the surface of infected HEK 293T cells. Following infection with recombinant AdC68, HEK 293T cells were analyzed by flow cytometry using a panel of mAbs targeting various epitopes in the pre- and post-F glycoproteins of RSV. The names and sites specificity of each mAb in the F glycoprotein of RSV are indicated. Motavizumab and Palivizumab, two clinically evaluated antibody drugs, recognize the identical epitope within site II. The X-axis indicates the PE fluorescence intensity while Y-axis denotes the number (counts) of the events. The percentage in each of the flow cytometry graph represents the percentage of PE-positive cells.
Article Snippet: Codon-optimized genes encoding the wild-type (WT) fusion glycoprotein (F) of RSV A2 strain (GenBank accession no: ACO83301.1), along with structure-based designer DS2, DS-Cav1, and SC-TM prefusion F glycoproteins – , were synthesized by GenScript Biotech Corporation, China.
Techniques: Bioprocessing, Recombinant, Plasmid Preparation, Expressing, Sequencing, Comparison, Infection, Flow Cytometry, Fluorescence