Antibody heavy- and light-chain variable regions were synthesized as “gBlocks” by Integrated DNA Technologies (www.idtdna.com) and subsequently cloned into IgG and IgK expression vectors as previously described56 (link). Equal ratios of heavy- and light-chain plasmids were co-transfected into 293F cells using FreeStyle MAX (Invitrogen) according to the manufacturer’s instructions. Protein G columns were used to purify IgG as previously described57 (link).
Simonich C.A., Doepker L., Ralph D., Williams J.A., Dhar A., Yaffe Z., Gentles L., Small C.T., Oliver B., Vigdorovich V., Mangala Prasad V., Nduati R., Sather D.N., Lee K.K., Matsen IV F.A, & Overbaugh J. (2019). Kappa chain maturation helps drive rapid development of an infant HIV-1 broadly neutralizing antibody lineage. Nature Communications, 10, 2190.
Other organizations :
Fred Hutch Cancer Center, University of Washington, Infectious Disease Research Institute, Center for Infectious Disease Research, University of Nairobi
Ratios of heavy- and light-chain plasmids co-transfected into 293F cells
dependent variables
Purified IgG
control variables
FreeStyle MAX (Invitrogen) transfection reagent used according to manufacturer's instructions
Protein G columns used to purify IgG as previously described
positive controls
Not explicitly mentioned
negative controls
Not explicitly mentioned
Annotations
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