1. Analysis of Env binding of Fluor-conjugated antibodies (A. Kliche)

2. -50 0 50 100 150 200 250 300 350 400 -50050100150200250 ConS gp140 #86 binding to A32 sTime RU Response A32 -50 0 50 100 150 200 250 300 -50050100150200250 Binding to ConS gp140 #86 sTime RU Response B12 - A32 B12-ALEXA488- A32 Req ConS 140 #86 - A32332 ConS 140 #86 - A32331 Req Ab binding ratio ConS 140 #86 + B12 - A32 274 0.825 ConS 140 #86 +B12-ALEXA488- A322500.755 ConS 140 A32 Ab A32 Control IgG

3. -50 0 50 100 150 200 250 300 350 -50050100150200250 ConS gp140 #86 sTime RU Response A32 -50 0 50 100 150 200 250 300 -50050100150200250 Binding to ConS gp140 #86 sTime RU Response HGN194 HGN194-ALEXA488 Req ConS 140 #86 - A32327 ConS 140 #86 - A32311 Req Ab binding ratio ConS 140 #86 + HGN194 - A322650.81 ConS140+HGN194-ALEXA488 - A322780.894 ConS 140 A32 Ab A32

4. -50 0 50 100 150 200 250 300 350 -50050100150200250 ConS 140 CFI Capture -50 0 50 100 150 200 250 -50050100150200250 HJ16 HJ16 Alexa Fluor 488 -50 0 50 100 150 200 250 300 350 -50050100150200250 ConS 140 CFI Capture -50 0 50 100 150 200 250 -50050100150200250300 HJ16 HJ16 Alexa Fluor 488 -50 0 50 100 150 200 250 300 -50050100150200250300350 ConS 140 CFI Capture -50 0 50 100 150 200 -1000100200300400 HJ16 HJ16 Alexa Fluor 488 Binding Response (RU) Time (s) ReplicateCapture Sample Binding Response (RU) Binding Antibody Binding Response (RU) Binding Ratio 1A32_ConS 140 CFI303HJ162080.686 2A32_ConS 140 CFI318HJ162410.758 3A32_ConS 140 CFI253HJ16180.60.714 1A32_ConS 140 CFI304HJ16 Alexa Fluor 48887.50.288 2A32_ConS 140 CFI316HJ16 Alexa Fluor 4881500.490 3A32_ConS 140 CFI215HJ16 Alexa Fluor 48892.30.431