(2010). luciferase transmission was reduced by >90% compared to the common transmission in six wells incubated with computer virus plus unfavorable control antibody. Unfavorable control: VP35-specific mAb. (E) Table showing initial mapping of the binding sites of the nine encouraging neutralizing mAbs. Left section: The promising mAbs (rows) were used as competitors to inhibit the binding of four reference mAbs (columns) to Jurkat cells expressing EBOV GP. Light-shaded cells show competition between a encouraging mAb and a reference mAb for binding. Right columns: The nine mAbs (rows) were tested for binding to previously explained Jurkat cell lines expressing EBOV GP variants with the indicated single amino-acid changes. Expression-normalized binding is usually shown as a percentage of binding to wild-type EBOV GP. Dark-shaded cells show significant loss of binding to a given GP variant. NIHMS1788146-product-2.tif (2.7M) GUID:?CD0CB891-3851-4D06-8A4E-B72EA1BBF78B 3: Physique S2. Cryo-EM local resolution estimation of antibody cocktail in complex with EBOV GP Trimer, related to Physique 2.The local resolution of the full map is estimated by Relion, showing the side view (A) and the top view Prim-O-glucosylcimifugin (B) of the mAbs and EBOV GP complex. (C) Zooming in on 1C3 variable region and its contacting region on GP. (D) Zooming in on 1C11 scFv and its contacting region on GP. NIHMS1788146-product-3.tif (4.7M) GUID:?7BE0A2FD-DA4B-44FC-9B25-9251E4B74A71 4: Physique S3. Broadly neutralizing antibody footprint comparison, related to Physique 3.The crystal structure of Ebola virus (EBOV) glycoprotein (GP) (PDB 5JQ3) is shown in surface representation with GP1 and GP2 colored light and dark gray, respectively. (A) The footprint of 1C11 is usually shown in blue, the footprint of 6D6 is usually shown in magenta, and their shared footprint is shown in purple. (B) The footprint of 1C11 is usually shown Prim-O-glucosylcimifugin in blue, the footprint of ADI-15878 is usually shown in yellow, and their shared footprint is shown in green. (C) A comparative sequence alignment to illustrate contact differences among 1C11 and the above four Prim-O-glucosylcimifugin broad neutralizing mAbs. Residues were labeled by colored triangles at the bottom to show the conservation (blue – conserved, pink – similar, yellow – non-conserved). NIHMS1788146-product-4.tif (3.0M) GUID:?81B1785A-5EB2-492C-808A-031C17B41198 5: Figure S4. Neutralization analysis of mAbs 1C3, related to Physique 4.(A) Point changes at four residues, P116, P123, P126, and D127 did not affect neutralization of rVSV-EBOV by 1C3. (B) GP residues in 1C3 contact sites that were changed in Physique 4B, ?,4C,4C, and S4A were labeled on the top view surface representation of EBOV GP trimer. The residues that did not affect neutralization were labeled in the left panel with circles, with colors corresponding to Prim-O-glucosylcimifugin Figure S4A. The residues Rabbit Polyclonal to Cyclin H that affected neutralization were labeled in the right panel with stars, with colors corresponding to Figure 4C. NIHMS1788146-product-5.tif (2.9M) GUID:?E9E5BB9A-A90F-4DFD-9198-5942C8A5A456 6: Figure S5. The binding of 1C3 Fab blocks three possible directions of NPC1-C access to EBOV GP, related to Physique 3 and ?and44.(A) Mab 1C3 Fab (light chain in yellow; heavy chain in orange) in complex with GP (GP1 in grey, GP2 in white), superimposed with NPC1 (dark blue) in complex with GP (PDB: Prim-O-glucosylcimifugin 5F1B). (B) The footprints of mAb 1C3 and NPC1-C on GP surface viewing from the top. Surface uniquely bound by 1C3 is in orange. Surface uniquely bound by NPC1 is in blue. The ten shared residues are in purple. NIHMS1788146-product-6.tif.