Cell Culture Conditions for Cytotoxicity Assays

SEM (55 (link)), Jurkat, CEM, MOLT-16 and Nalm-6 cells (DSMZ) were cultured in RPMI 1640 GlutaMax-I medium (Thermo Fisher Scientific) supplemented with 10% fetal calf serum (FCS; Thermo Fisher Scientific), 100 U/mL penicillin and 100 µg/mL streptomycin (Thermo Fisher Scientific) (R10+). BHK-CD16a (FcγRIIIa V158) cells were cultured in R10+ medium supplemented with 10 μmol/l methotrexate (Sigma-Aldrich) and 500 μg/ml geneticin (Thermo Fisher Scientific) as described before (53 (link), 54 (link), 56 (link)). CHO-CD32a (FcγRIIa H131) cells were cultivated in DMEM medium (Thermo Fisher Scientific) supplemented with 10% FCS, 100 U/ml penicillin and 100 µg/ml streptomycin (57 (link)). Medium was supplemented with 1% NEAA (Thermo Fisher Scientific), 1% sodiumpyruvat (Thermo Fisher Scientific) and 500 µg/ml geneticin. Chinese hamster ovary cells (CHO-S, Thermo Fisher Scientific) were cultured in serum-free CD-CHO medium (Thermo Fisher Scientific) containing 1% HT supplement (Thermo Fisher Scientific) and 2 mM GlutaMax (Thermo Fisher Scientific). After transfection CHO-S cells were cultured in CD OptiCHO medium (Thermo Fisher Scientific) containing 1% HT supplement, 2 mM GlutaMax and 0.1% Pluronic F-68 (Thermo Fisher Scientific).

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Gehlert C.L., Rahmati P., Boje A.S., Winterberg D., Krohn S., Theocharis T., Cappuzzello E., Lux A., Nimmerjahn F., Ludwig R.J., Lustig M., Rösner T., Valerius T., Schewe D.M., Kellner C., Klausz K, & Peipp M. (2022). Dual Fc optimization to increase the cytotoxic activity of a CD19-targeting antibody. Frontiers in Immunology, 13, 957874.

Publication 2022

RPMI 1640 Glutamax-I medium penicillin streptomycin methotrexate geneticin DMEM medium NEAA sodiumpyruvat CHO-S CD-CHO medium GlutaMax CD OptiCHO medium HT supplement Pluronic F-68

Cd32a Cells Cho cells Cultured medium Fcγriia Geneticin Methotrexate Molt Penicillin Pluronic f 68 Serum Streptomycin Supplement Transfection

Corresponding Organization : Kiel University

Other organizations : University Hospital Schleswig-Holstein, University of Lübeck, University of Padua, Friedrich-Alexander-Universität Erlangen-Nürnberg, Otto-von-Guericke University Magdeburg, Ludwig-Maximilians-Universität München

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Variable analysis

independent variables

Cell lines used: SEM (55), Jurkat, CEM, MOLT-16, Nalm-6, BHK-CD16a (FcγRIIIa V158), CHO-CD32a (FcγRIIa H131), and CHO-S

dependent variables

Not explicitly mentioned

control variables

Culture medium: RPMI 1640 Glutamax-I supplemented with 10% fetal calf serum (FCS), 100 U/mL penicillin, and 100 µg/mL streptomycin (R10+)
BHK-CD16a (FcγRIIIa V158) cells were cultured in R10+ medium supplemented with 10 μmol/l methotrexate and 500 μg/ml geneticin
CHO-CD32a (FcγRIIa H131) cells were cultivated in DMEM medium supplemented with 10% FCS, 100 U/ml penicillin, 100 µg/ml streptomycin, 1% NEAA, 1% sodiumpyruvat, and 500 µg/ml geneticin
CHO-S cells were cultured in serum-free CD-CHO medium containing 1% HT supplement and 2 mM GlutaMax, and after transfection they were cultured in CD OptiCHO medium containing 1% HT supplement, 2 mM GlutaMax and 0.1% Pluronic F-68

controls

No positive or negative controls were explicitly mentioned in the provided information.

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