non competing anti gp38 antibodies  (Teknova)

Journal: Cell reports

Article Title: Crimean-Congo hemorrhagic fever survivors elicit protective non-neutralizing antibodies that target 11 overlapping regions on glycoprotein GP38

doi: 10.1016/j.celrep.2024.114502

Figure Lengend Snippet: (A) Matrix of percent sequence identity of GP38 amino acid residues (AA) across six CCHFV isolates. (B) Single concentration BLI binding analysis of 188 antibodies to the six rGP38 proteins as a whole panel (top) and broken down by bin (bottom). Shades of green represent the number of rGP38 proteins bound by a single antibody (from 0 in gray to 6 in darkest green). Total number of mAbs is indicated in the circular diagram and total mAbs from each bin are indicated above the bar graph. (C) Carterra system HT-SPR binding analysis of six lead antibody candidates binding to six rGP38 proteins. The highest binding affinities are in dark green and the lowest binding affinities are in white. Calculated K D values appear in each rectangle of the heatmap; for samples that were off-rate limited, K D values are denoted as < the calculated K D . The one interaction for which a curve could not be fit is denoted as P.F.

Article Snippet: Cells that bound the non-competing anti-GP38 antibodies were sorted and plated on complete minimal media glucose agar plates minus tryptophan (Teknova).

Techniques: Sequencing, Concentration Assay, Binding Assay

Journal: Cell reports

Article Title: Crimean-Congo hemorrhagic fever survivors elicit protective non-neutralizing antibodies that target 11 overlapping regions on glycoprotein GP38

doi: 10.1016/j.celrep.2024.114502

Figure Lengend Snippet: (A) Yeast-based mapping strategy of select antibodies to identify critical binding residues on GP38. The percentage of antibody binding retained by each GP38 variant is colored according to the key. Critical residues are defined as mutations that led to a binding disruption of 75% or more and are colored by the assigned antigenic site. (B) Yeast-based critical residues mapped on the surface of GP38: bin I (blue, residues Val385, Pro388), bin II (green, residues Gly371, Leu374, Ile375, Lys404, Lys488, Leu499), bin III (yellow, residues Ser428-Ala429, Asp444-Asp446, Lys474-Leu475, Asp477), bin IV (orange, residues Ile253-Leu255, Leu257, Lys262, Gly266, Glu277, Glu281), bin V (red, residues Glu285, Arg289, Gly292). (C) Composite structure of GP38 bound with representative antibodies. GP38 is shown as a rainbow ribbon and Fabs as molecular surfaces. Heavy chains are colored to represent the five non-overlapping bins, and light chains are white. Black dashed lines highlight the vertical alignment of Fabs along one plane (left) and the opposing binding directions to another plane (right).

Article Snippet: Cells that bound the non-competing anti-GP38 antibodies were sorted and plated on complete minimal media glucose agar plates minus tryptophan (Teknova).

Techniques: Binding Assay, Variant Assay, Disruption

Journal: Cell reports

Article Title: Crimean-Congo hemorrhagic fever survivors elicit protective non-neutralizing antibodies that target 11 overlapping regions on glycoprotein GP38

doi: 10.1016/j.celrep.2024.114502

Figure Lengend Snippet: (A) Crystal structure of GP38 bound with ADI-46143 (bin I, blue) with heavy-chain interactions (top) and light-chain interactions (bottom). (B) Cryo-EM structure of GP38 bound with ADI-58048 (bin II, green, left) and ADI-46152 (bin IV+V, red, right). Heavy-chain interactions (top left, top right) and light-chain interactions (bottom left, bottom right) are shown in the insets. (C) Crystal structure of GP38 bound with c13G8 (bin IV+V, red) with heavy-chain interactions (top) and light-chain interactions (bottom). For all panels, heavy chains are colored, light chains are gray, polar interactions are indicated by black dashed lines, and GP38 residues are labeled in white text with a black outline.

Article Snippet: Cells that bound the non-competing anti-GP38 antibodies were sorted and plated on complete minimal media glucose agar plates minus tryptophan (Teknova).

Techniques: Cryo-EM Sample Prep, Labeling

Journal: Cell reports

Article Title: Crimean-Congo hemorrhagic fever survivors elicit protective non-neutralizing antibodies that target 11 overlapping regions on glycoprotein GP38

doi: 10.1016/j.celrep.2024.114502

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: Cells that bound the non-competing anti-GP38 antibodies were sorted and plated on complete minimal media glucose agar plates minus tryptophan (Teknova).

Techniques: Derivative Assay, Virus, Recombinant, Protease Inhibitor, Reverse Transcription, Membrane, Cell Isolation, Mutagenesis, Expressing, Plasmid Preparation, Software, Affinity Column, Sequencing, Transfection, Electron Microscopy