1
Figure S1
Figure S1. PAG frequency analysis for mAbs 2F5, Z13e1, 2909, and b12 bound to unliganded and CD4-liganded HIV-1 MN, BaL and SF162 strains. Arrows and lines as in figure 4.
Figure S2
Figure S2. PAG frequency analysis for mAbs 2F5, Z13e1 and 2909 bound to the unliganded and CD4-liganded SIV short-tailed. Arrows and lines as in figure 4.
Figure S3
Figure S3. PAG frequency analysis for Fab 4E10 inhibition assay. Arrows and lines as in figure 4.
Figure S4
Figure S4. PAG frequency analysis for the mAb 4E10 and mAb 2909 washout assay. Arrows and lines as in figure 4.
Figure S5
Figure S5. PAG frequency analysis for mAb 2F5 washout assay. Arrows and lines as in figure 4.
Figure S6
Figure S6. PAG frequency analysis for mAb Z13e1 washout assay. Arrows and lines as in figure 4.
Table S1
Biochemical estimates of Env spikes per virion
Virions / Gag/Envratio / Trimers per virion
SIV239/251 tail/SupT1-CCR5 / 13-19 : 1 / 26-38
SIVmac239/CEMX 174(T1) / 60 : 1 / 7-14
SIV*4E10/SupT1-CCR5 / 9-14 : 1 / 34-58
HIV-1 (MN) CL.4/SupT1-R / 56-67 : 1 / 8-10
HIV-1SF162/SupT1-CCR5 / 81-160 : 1 / 3-6
Table S2. Standard PAG binding assay statistical data
Reactants / Mean / SD / SE / Range / Median / NHIV-1 MN + mAb 4E10 / 3.63 / 2.87 / 0.23 / 0-12 / 3 / 159
HIV-1 MN + sCD4 + mAb 4E10 / 10.23 / 6.28 / 0.46 / 0-34 / 10 / 183
HIV-1 MN + 2F5 / 5.41 / 3.73 / 0.45 / 0-16 / 6 / 70
HIV-1 MN + sCD4 + 2F5 / 8.23 / 5.76 / 0.65 / 0-26 / 8 / 78
HIV-1 MN + mAb Z13e1 / 3.83 / 3.03 / 0.21 / 0-16 / 3 / 207
HIV-1 MN + sCD4 + mAb Z13e1 / 8.31 / 6.26 / 0.49 / 0-46 / 8 / 163
HIV-1 MN + mAb b12 / 9.27 / 6.28 / 0.65 / 0-29 / 8 / 93
HIV-1 MN + sCD4 + mAb b12 / 1.53 / 1.47 / 0.14 / 0-5 / 1 / 107
HIV-1 MN + mAb 2909 / 0.62 / 1.00 / 0.09 / 0-5 / 0 / 121
HIV-1 MN + sCD4 + mAb 2909 / 0.39 / 0.79 / 0.07 / 0-4 / 0 / 115
HIV-1 BaL + mAb 4E10 / 2.93 / 3.41 / 0.32 / 0-28 / 2 / 213
HIV-1 BaL + sCD4 + mAb 4E10 / 4.80 / 3.98 / 0.23 / 0-18 / 4 / 156
HIV-1 SF162 + mAb 4E10 / 2.10 / 2.22 / 0.14 / 0-11 / 1 / 263
HIV-1 SF162 + sCD4 + mAb 4E10 / 4.78 / 3.86 / 0.33 / 0-24 / 4 / 139
SIV short-tailed + mAb 4E10 / 3.10 / 3.06 / 0.18 / 0-18 / 3 / 277
SIV short-tailed + sCD4 + mAb 4E10 / 4.55 / 4.00 / 0.24 / 0-28 / 4 / 276
SIV short-tailed + mAb 2F5 / 2.37 / 2.16 / 0.18 / 0-10 / 2 / 139
SIV short-tailed + sCD4 + mAb 2F5 / 1.35 / 1.91 / 0.20 / 0-12 / 1 / 91
SIV short-tailed + mAb Z13e1 / 2.35 / 1.91 / 0.16 / 0-9 / 2 / 150
SIV short-tailed + sCD4 + mAb Z13e1 / 1.63 / 1.66 / 0.14 / 0-8 / 1 / 148
SIVmac239 + mAb 4E10 / 1.57 / 1.64 / 0.14 / 0-6 / 1 / 128
SIVmac239 + sCD4 + mAb 4E10 / 1.29 / 1.27 / 0.13 / 0-7 / 1 / 90
SIV-4E10 + mAb 4E10 / 7.91 / 6.48 / 0.56 / 0-34 / 6 / 134
SIV-4E10 + sCD4 + mAb 4E10 / 19.02 / 9.85 / 0.55 / 0-58 / 20 / 316
SIV short-tailed + mAb 2909 / 0.36 / 0.69 / 0.08 / 0-3 / 0 / 75
SIV short-tailed + sCD4 + mAb 2909 / 0.34 / 0.75 / 0.09 / 0-4 / 0 / 68
1
Table S3. PAG Fab inhibition assay statistical data
Reactants / Mean / SD / SE / Range / Median / NHIV-1 + 4E10 normalized / 3.68 / 2.75 / 0.22 / 0-14 / 4 / 154
HIV-1 + Fab 4E10 + 4E10 / 1.13 / 2.01 / 0.18 / 0-11 / 0 / 132
HIV-1 + sCD4 + 4E10 normalized / 8.74 / 5.34 / 0.61 / 0-25 / 8 / 76
HIV-1 + sCD4 + Fab 4E10 + 4E10 / 1.85 / 2.19 / 0.18 / 0-10 / 1 / 142
SIV-4E10 + 4E10 normalized / 10.16 / 7.64 / 0.90 / 0-32 / 11 / 73
SIV-4E10 + Fab 4E10 + 4E10 / 2.57 / 2.40 / 0.23 / 0-11 / 2 / 108
SIV-4E10 + sCD4 + 4E10 normalized / 18.43 / 11.48 / 1.39 / 0-48 / 19 / 68
SIV-4E10 + sCD4 + Fab 4E10 + 4E10 / 2.28 / 3.16 / 0.30 / 0-24 / 2 / 109
SIV short-tailed + 4E10 normalized / 2.69 / 2.73 / 0.27 / 0-18 / 2 / 100
SIV short-tailed + Fab 4E10 + 4E10 / 1.12 / 1.49 / 0.12 / 0-8 / 1 / 149
SIV Microvesicles + 4E10 normalized / 2.60 / 5.37 / 0.58 / 0-34 / 0 / 87
SIV microvesicles + Fab 4E10 + 4E10 / 0.97 / 1.79 / 0.22 / 0-9 / 0 / 68
Table S4. PAG washout assay statistical data
Reactants / Mean / SD / SE / N / MedianNo wash / Washout / No wash / Washout / No wash / Washout / No wash / Washout / No wash / Washout
HIV-1 + mAb 4E10 / 3.95 / 3.94 / 2.82 / 3.13 / 0.26 / 0.27 / 116 / 133 / 4 / 4
SIV-4E10 + mAb 4E10 / 7.08 / 7.82 / 5.14 / 4.62 / 0.33 / 0.34 / 246 / 186 / 6 / 7
SIV short-tailed + mAb 4E10 / 2.53 / 2.87 / 2.24 / 2.57 / 0.24 / 0.29 / 90 / 77 / 2 / 2
Microvesicles + mAb 4E10 / 2.04 / 1.56 / 5.85 / 4.77 / 0.54 / 0.43 / 118 / 124 / 0 / 0
SIV short-tailed + mAb 2909 / 0.45 / 0.33 / 0.84 / 0.72 / 0.05 / 0.04 / 275 / 294 / 0 / 0
HIV-1 + mAb 2F5 / 3.72 / 3.08 / 3.07 / 2.38 / 0.21 / 0.14 / 208 / 272 / 3 / 3
SIV short-tailed + mAb 2F5 / 1.78 / 0.91 / 1.82 / 1.15 / 0.16 / 0.06 / 133 / 322 / 1 / 1
Microvesicles + mAb 2F5 / 1.07 / 0.43 / 1.90 / 1.26 / 0.16 / 0.09 / 149 / 183 / 0 / 0
HIV-1 + mAb Z13E1 / 2.93 / 3.52 / 2.45 / 3.03 / 0.14 / 0.18 / 296 / 288 / 3 / 3
SIV short-tailed + mAb Z13E1 / 1.87 / 1.86 / 1.79 / 1.75 / 0.13 / 0.20 / 182 / 79 / 2 / 2
Microvesicles + mAb Z13E1 / 1.46 / 1.52 / 2.30 / 2.14 / 0.14 / 0.15 / 285 / 199 / 1 / 1
Table S5. PAG kinetics of mAb 4E10 binding to HIV-1 MN
Kinetics of mAb 4E10 binding / Time (min) / Mean / SD / SE / Range / Median / NUnliganded HIV-1 + mAb 4E10 / 15 / 4.37 / 3.04 / 0.30 / 0-13 / 4 / 101
30 / 5.50 / 3.43 / 0.36 / 0-18 / 5 / 92
70 / 6.52 / 5.30 / 0.38 / 0-27 / 6 / 196
600 / 7.70 / 4.59 / 0.29 / 0-24 / 7.5 / 248
1080 / 8.94 / 4.09 / 0.38 / 0-22 / 9 / 115
CD4 co-liganded HIV-1 with mAb 4E10 / 15 / 4.25 / 3.59 / 0.34 / 0-17 / 4 / 112
30 / 9.68 / 5.21 / 0.67 / 0-24 / 10 / 160
70 / 10.93 / 6.82 / 0.62 / 0-30 / 10 / 121
600 / 10.09 / 5.88 / 0.49 / 0-27 / 10 / 141
1080 / 10.53 / 5.35 / 0.37 / 0-28 / 10 / 212
CD4 pre-liganded HIV-1 + mAb 4E10 / 15 / 9.69 / 5.98 / 0.60 / 0-25 / 10 / 98
30 / 9.05 / 5.85 / 0.57 / 0-28 / 8 / 105
70 / 11.25 / 6.83 / 0.56 / 0-37 / 11 / 151
600 / 10.44 / 5.42 / 0.45 / 0-28 / 10 / 143
1080 / 9.73 / 5.04 / 0.36 / 0-21 / 10 / 192
Unliganded HIV-1 gp120 shedding control (2G12 binding) / 15 / 8.48 / 5.64 / 0.46 / 0-22 / 8 / 149
30 / 8.60 / 6.47 / 0.49 / 0-34 / 7 / 176
70 / 8.14 / 4.67 / 0.30 / 0-21 / 8 / 242
190 / 7.78 / 4.97 / 0.42 / 0-24 / 8 / 140
600 / 7.86 / 4.71 / 0.23 / 0-28 / 8 / 406
1080 / 8.72 / 5.81 / 0.31 / 0-31 / 8 / 362
CD4 pre-liganded HIV-1 gp120 shedding control (2G12 binding) / 15 / 7.08 / 4.48 / 0.46 / 0-17 / 6.5 / 96
30 / 8.15 / 5.93 / 0.42 / 0-26 / 8 / 203
70 / 8.22 / 5.08 / 0.54 / 0-22 / 8.5 / 90
190 / 7.77 / 5.01 / 0.35 / 0-26 / 7 / 201
600 / 4.21 / 4.11 / 0.23 / 0-20 / 3 / 308
1080 / 4.00 / 3.35 / 0.19 / 0-17 / 3 / 321