high throughput screening cytometer  (Sartorius AG)

Journal: The Journal of Clinical Investigation

Article Title: Human antibody targeting Crimean-Congo hemorrhagic fever virus glycoprotein 38 protects mice against heterologous virus challenge

doi: 10.1172/JCI191440

Figure Lengend Snippet: PreGn binding antibodies were titrated and further assessed for their capacity to bind preGn-expressing cells. mAbs were serially titrated and allowed to incubate on preGn transiently expressing cells before washing and adding PE-conjugated secondary antibody. Secondary antibody was washed, and cells were resuspended for analysis on an iQue flow cytometer. Binding to transfected cells was observed relative to untransfected cells, and data are presented as mean values ± SD and represent duplicate values from each of 3 independent experiments. IC 50 values were calculated using a 4-parameter sigmoidal nonlinear fit in Prism software version 9 (GraphPad). CCHF-245 was used as a negative control.

Article Snippet: After 7 days, LCL supernatants were screened for the presence of CCHFV Gc-, Gn-, or GP38-recognizing antibodies using IbAr10200 strain M segment–transfected Expi293F cells on the Intellicyt iQue high-throughput flow cytometer as described below.

Techniques: Binding Assay, Expressing, Flow Cytometry, Transfection, Software, Negative Control

Journal: The Journal of Clinical Investigation

Article Title: Human antibody targeting Crimean-Congo hemorrhagic fever virus glycoprotein 38 protects mice against heterologous virus challenge

doi: 10.1172/JCI191440

Figure Lengend Snippet: PreGn binding antibodies were added to preGn-expressing cells at saturating concentrations (20 μg/mL) for 30 minutes. Alexa Fluor 647–conjugated mouse or human antibodies were then added to cells at 5 μg/mL without a prior wash step. Cells were washed and immediately processed on an iQue flow cytometer. The values shown are the percentage of binding that occurred during competition compared with noncompeted binding of the mAb. This value was normalized to 100%. The values are also indicated by the dotted lines; above 40% indicated no competition, between 40% and 20% indicated partial competition, and below 20% indicated competition. Values shown are from 2 independent experiments with 3 replicates. Data represent mean ± SD.

Article Snippet: After 7 days, LCL supernatants were screened for the presence of CCHFV Gc-, Gn-, or GP38-recognizing antibodies using IbAr10200 strain M segment–transfected Expi293F cells on the Intellicyt iQue high-throughput flow cytometer as described below.

Techniques: Binding Assay, Expressing, Flow Cytometry