Purification of anti-PfCSP human monoclonal antibodies
The isolation of PfCSP hmAbs used in this study was previously reported.8 (link)–12 (link) All hmAbs were expressed in IgG1 heavy and light chain plasmids using the ExpiFectamine™ 293 Transfection Kit (Thermo Fisher Scientific) and incubated at 37°C, 8% CO2 for 6 days. Supernatants were harvested and purified using rProtein A Sepharose Fast Flow resin (GE Healthcare) or protein G chromatography cartridge (Thermo Fisher Scientific). Buffer was exchanged with 1X PBS before being concentrated using Amicon Centrifugal Filters (Millipore). The concentration of purified antibodies was determined using the Nanodrop spectrophotometer (Thermo Fisher Scientific).
Aguirre-Botero M.C., Wang L.T., Formaglio P., Aliprandini E., Thiberge J.M., Schön A., Flores-Garcia Y., Mathis-Torres S., Flynn B.J., da Silva Pereira L., Le Duff Y., Hurley M., Nacer A., Bowyer P.W., Zavala F., Idris A.H., Francica J.R., Seder R.A, & Amino R. (2023). Cytotoxicity of human antibodies targeting the circumsporozoite protein is amplified by 3D substrate and correlates with protection. Cell reports, 42(7), 112681.
Other organizations :
Université Paris Cité, Institute of Infection and Immunity, Sorbonne Paris Cité, National Institutes of Health, Johns Hopkins University, National Institute of Malaria Research, National Institute for Biological Standards and Control
Expression of hmAbs in IgG1 heavy and light chain plasmids
Incubation at 37°C, 8% CO2 for 6 days
dependent variables
Antibody expression levels
Antibody purification yield
control variables
Use of the ExpiFectamine™ 293 Transfection Kit
Purification using rProtein A Sepharose Fast Flow resin or protein G chromatography cartridge
Buffer exchange with 1X PBS
Concentration using Amicon Centrifugal Filters
Antibody concentration determination using Nanodrop spectrophotometer
controls
No positive or negative controls were explicitly mentioned in the provided information.
Annotations
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