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Journal: Immunity
Article Title: Multifunctional Pan-ebolavirus Antibody Recognizes a Site of Broad Vulnerability on the Ebolavirus Glycoprotein
doi: 10.1016/j.immuni.2018.06.018
Figure Lengend Snippet: EBOV-515 and -520 Target Both Intact GP and Cleaved GP CL Intermediate to Neutralize the Virus (A) Binding curves for EBOV-515 or -520 using Jurkat-EBOV GP or Jurkat-EBOV GP CL . Fluorescently labeled mAbs were incubated with cells, and binding was assessed by flow cytometric analysis. (B) Neutralization curves for EBOV-515 or -520 or control mAbs 13C6 or BDBV317 using rVSV/EBOV-GP or rVSV/EBOV-GP CL . (C) Capacity of mAbs to inhibit NPC1-C binding to GP CL . mAbs were incubated with Jurkat-EBOV GP CL , then with purified NPC1-C tagged with FLAG-epitope. Complexes were detected with anti-FLAG Abs by flow cytometry. (D) Competition binding of EBOV-520 with RBS-specific mAbs MR72 or MR78 was assessed using Jurkat-EBOV GP CL . mAb binding was analyzed by flow cytometry. Numbers indicate the percent binding of the second fluorescently labeled mAb (2) in the presence of the first unlabeled mAb (1), compared to binding of second labeled mAb alone (dotted line). (E) Binding curves of fluorescently labeled mAb EBOV-515 or -520 to Jurkat-EBOV GP in the presence of a fixed concentration of unlabeled mAb EBOV-437 or -442. Mean ± SD of triplicates are shown, and data in (A) and (C)–(E) are representative of two independent experiments. Data in (B) represent one experiment. See also Figure S4 .
Article Snippet: Recombinant NPC1 protein, His/FLAG-tagged , Creative BioMart , Cat# NPC1-1339H.
Techniques: Virus, Binding Assay, Labeling, Incubation, Neutralization, Control, Purification, FLAG-tag, Flow Cytometry, Concentration Assay