The monoclonal antibodies (mAbs) 17b [60 (link)], 35O22 [61 (link)], 447-52D [62 (link)], F105 [63 (link)], F240 [64 (link)], PGT121 [65 (link)], and Z13e1 [66 (link)] were obtained from the National Institutes of Health (NIH) HIV Reagent Program (HRP). Anti-Env mAbs 2F5, 2G12, 4E10, PG9, and PG16 were purchased from Polymun (Vienna, Austria). Anti-p24 mAb BC1071 and sheep polyclonal antibodies anti-p24 (D7320) and anti-gp120 (D7324) were purchased from Aalto Bio Reagents (Dublin, Ireland). Anti-GAPDH (14C10) rabbit mAb was purchased from Cell Signaling Technology (Danvers, MA, USA).
Heavy- and light-chain expression plasmids for 10E8, PGT151, and CAP256-VRC26.06 (synthesized from published protein sequences [67 (link),68 (link),69 (link)], respectively) were kindly obtained from Pantelis Poumbourios (Burnet Institute, Melbourne, Australia), and VRC01 expression plasmids were obtained from the NIH HRP [70 (link)]. Antibody heavy and light chains were coexpressed in Expi293F cells and purified using Protein G Agarose Fast Flow (MilliporeSigma) according to the manufacturer’s instructions.
Heavy- and light-chain expression plasmids for 10E8, PGT151, and CAP256-VRC26.06 (synthesized from published protein sequences [67 (link),68 (link),69 (link)], respectively) were kindly obtained from Pantelis Poumbourios (Burnet Institute, Melbourne, Australia), and VRC01 expression plasmids were obtained from the NIH HRP [70 (link)]. Antibody heavy and light chains were coexpressed in Expi293F cells and purified using Protein G Agarose Fast Flow (MilliporeSigma) according to the manufacturer’s instructions.
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