Figure 5.

Analysis of the affinities of 447–52D and HGN194 to the individual members of the Env/V3 model library. 24 h post induction of the Env/V3 stable HEK293 cell lines with doxycycline (1 µg/ml), these cells were stained with mAbs 447–52D or HGN194 until equilibrium binding was reached. Relative MFI values of bound antibodies in relation to the eGFP signal (MFI APC/ MFI eGFP) to normalize for Env expression levels are shown (mean values of 3 independent experiments). A and C: Relative MFIs (MFI APC/ MFI eGFP) of HEK293 cell lines stained with 2.7 nM 447–52D (A) and 6.7 nM HGN194 (C) (the same concentrations as used in all sorting experiments, see below) are shown to illustrate the individual binding pattern of the 5 Env/V3 chimeras to the respective antibody. B and D: flow cytometry titration using serial dilutions of 447–52D (B) and HGN194 (D).