Affinofile profiling reveals that resistance to broadly neutralizing antibodies (BNAbs) also results in reduced entry efficiency. (A-C) N160K and N279/280A mutations were engineered into a random sample of 12 subtype A-D Envs. The resultant (PG9/PG16)R and (VRC01)R resistant Envs were assayed for CD4 and CCR5 usage efficiency along with their parental BNAb sensitive Envs. GGR Affinofile profiling was performed as previously described. 2-D contour plots of the averaged infectivity profiles for (A) WT, (B) (PG9/PG16)R, and (C) (VRC01)R Envs are shown. The infectivity profile for the individual Envs are shown in supplementary Figure S5. Axes and color-codes are identical to previous contour plots. (D-E) The median values and interquartile ranges of the Mean infectivity (M) are shown for (PG9/PG16)R or (VRC01)R resistant Envs compared to their WT counterparts. P values calculated via a non-parametric paired t-test.