Journal: Emerging Microbes & Infections

Article Title: A pan-orthoebolavirus neutralizing antibody encoded by mRNA effectively prevents virus infection

doi: 10.1080/22221751.2024.2432366

Figure Lengend Snippet: The mAb 2G1 exhibited a broad anti-orthoebolaviruses activity. (A) Recognition of 2G1 to the full length of orthoebolavirus GPs displayed on the cell surface. Data represent mean ± SD of three replicates from one representative experiment. An FITC-labeled irrelevant mAb was used as the control. (B) Binding kinetics of 2G1 to EBOV GPΔTM or GPΔMuc determined by surface plasmon resonance. Five representative curves from one representative test were fitted to compute the kinetic constants. (C) Binding activities of 2G1 to EBOV GPΔTM, GPΔMuc, and GPcl under various pH conditions tested by ELISA. (D) Neutralization capacities of 2G1, FVM04, and mAb114 against pseudotyped rHIV-EBOV (Makona), – EBOV (Mayinga), – SUDV, – BDBV, – TAFV, and – RESTV. Data are presented as the mean ± SD of three replicates from one representative experiment. (E) The amino acid divergence between the original and inferred genes of 2G1. (F) The binding abilities of the wild-type and inferred germline cross-pairing forms of 2G1 to EBOV GPΔMuc, tested by ELISA. (G) The binding curves of HCDR mutants of 2G1 to EBOV GPΔMuc.

Article Snippet: Recombinant EBOV GPΔTM was purchased from Sino Biological Inc. (Beijing, China; Cat No. 40442-V08H4).

Techniques: Activity Assay, Labeling, Control, Binding Assay, SPR Assay, Enzyme-linked Immunosorbent Assay, Neutralization

Journal: Emerging Microbes & Infections

Article Title: A pan-orthoebolavirus neutralizing antibody encoded by mRNA effectively prevents virus infection

doi: 10.1080/22221751.2024.2432366

Figure Lengend Snippet: 2G1 recognized the hydrophobic pocket beneath the N-terminal tail of GP2 and inhibited GP proteolysis. (A) Front and top views of the cryo-EM structure of the EBOV GPΔMuc/2G1 Fab complex. Molecules are shown as surface representations. The GP1/2 subunits of three protomers A – C of GPΔMuc are colored in pale/blue violet, light/dodger blue, and light/hot pink, respectively. The VH of 2G1 is colored in lime green, and the VL is colored in goldenrod. (B) Superimposition of the 2G1, KZ52 (PDB ID: 5HJ3), and ADI-15878 (PDB ID: 6EA7) variable regions onto EBOV GPΔMuc. Molecules are shown as surface representations. 2G1, KZ52, and ADI-15878 were colored goldenrod, light sea green, and lime green, respectively. (C) Footprints of 2G1 or ADI-15878 on the EBOV GPΔMuc. The buried areas of 2G1 and ADI-15878 are indicated by goldenrod and lime green dotted lines, respectively. The paratope residues of 2G1 are shown as sticks. (D) Magnified view of the interface between 2G1 and EBOV GPΔMuc. EBOV GPΔMuc is represented as a molecular surface, the interface residues are shown as stick representations, and H-bonds are indicated by dotted black lines. GP1 (protomer A), GP2 (protomer A), GP2 (protomer B), 2G1 HCDR, 2G1 LCDR, oxygen atoms, and nitrogen atoms are colored pale violet, blue violet, dodger blue, lime green, goldenrod, red, and blue, respectively. (E) Relative binding percentage (%WT) of non-competing 2G1 and 4F1 to surface-anchored EBOV GP mutants. Data represent the means of three replicates of one representative experiment. Minor and pivotal epitopes of 2G1 are indicated in orange and dodger blue, respectively. (F) Conservation of critical 2G1 epitopes among filovirus GPs. (G) Antibodies blocking the binding of NPC1-C to GPcl. (H) Ability of antibodies or Fabs to inhibit the cleavage of EBOV GPΔMuc by thermolysin. THL, thermolysin; IMF, intermediate mass fragment.

Article Snippet: Recombinant EBOV GPΔTM was purchased from Sino Biological Inc. (Beijing, China; Cat No. 40442-V08H4).

Techniques: Cryo-EM Sample Prep, Binding Assay, Blocking Assay

Journal: Emerging Microbes & Infections

Article Title: A pan-orthoebolavirus neutralizing antibody encoded by mRNA effectively prevents virus infection

doi: 10.1080/22221751.2024.2432366

Figure Lengend Snippet: mRNA-2G1-LNP potently inhibited the pseudotyped EBOV and SUDV infection. (A) The neutralization ability of mRNA-2G1-LNP against rHIV-EBOV. (B and C) The in vivo protection efficacy of mRNA-2G1-LNP against rHIV-EBOV in mice. The normalized bioluminescence intensity (p/s/cm2/sr) of images captured four days post-infection (B) was calculated and compared to the PBS group (C). (D – F) In vitro neutralization ability (D) and in vivo protective efficacy (E and F) of mRNA-2G1-LNP against rHIV-SUDV. (G) Serum antibody concentration. BALB/c mice ( n = 5) were administered a single dose of mRNA-2G1-LNP, 2G1 antibody, or an equal volume of empty LNP. Antibody concentrations at the indicated times were determined using quantitative ELISA. (I – L) Comparison of biochemical criteria in the serum three days post-administration between the mRNA-2G1-LNP (1 mg/kg) and PBS groups. ALT, alanine aminotransferase; AST, aspartate aminotransferase; LDH, lactate dehydrogenase; CR, creatinine.

Article Snippet: Recombinant EBOV GPΔTM was purchased from Sino Biological Inc. (Beijing, China; Cat No. 40442-V08H4).

Techniques: Infection, Neutralization, In Vivo, In Vitro, Concentration Assay, Enzyme-linked Immunosorbent Assay, Comparison