Recombinant Anti-HIV-1 nef VHH Single Domain Antibody (PNBL-022) (CAT#: PNBL-022)

Figure 1 Structure of the tripartite SH3B₆–Nef–sdAb19 complex.

Binding of the camelid antibody sdAb19 to HIV-1 Nef showed a dissociation constant of 39 nM.

Lülf, S., Matz, J., Rouyez, M. C., Järviluoma, A., Saksela, K., Benichou, S., & Geyer, M. (2014). Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 11(1), 24.

Figure 2 Functional analyses of Nef–sdAb19 interactions.

Mutation of D60R in sdAb19 gradually attenuated binding to Nef as determined by ITC measurements.

Lülf, S., Matz, J., Rouyez, M. C., Järviluoma, A., Saksela, K., Benichou, S., & Geyer, M. (2014). Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 11(1), 24.

Figure 3 Functional analyses of Nef–sdAb19 interactions.

Mutation of G102R/S103E in sdAb19 gradually attenuated binding to Nef as determined by ITC measurements.

Lülf, S., Matz, J., Rouyez, M. C., Järviluoma, A., Saksela, K., Benichou, S., & Geyer, M. (2014). Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 11(1), 24.

Figure 4 Functional analyses of Nef–sdAb19 interactions.

Mutation of the combined triple mutant D60R/G102R/S103E in sdAb19 gradually attenuated binding to Nef as determined by ITC measurements.

Lülf, S., Matz, J., Rouyez, M. C., Järviluoma, A., Saksela, K., Benichou, S., & Geyer, M. (2014). Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 11(1), 24.

Figure 5 Functional analyses of Nef–sdAb19 interactions.

Mutant sdAb19 proteins abrogate the inhibitory effect of sdAb19 on CD4 internalization. HeLa-CD4 cells were transfected with plasmids for expression of either Nef-GFP or GFP in combination with the plasmid for expression of wild-type or mutated sdAb19.

Lülf, S., Matz, J., Rouyez, M. C., Järviluoma, A., Saksela, K., Benichou, S., & Geyer, M. (2014). Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 11(1), 24.

Figure 6 Neffin binding to Nef results in a 2:2 complex formation.

Isothermal titration calorimetry confirms tight binding between Nef and Neffin exhibiting a Kd of 1.6 nM.

Lülf, S., Matz, J., Rouyez, M. C., Järviluoma, A., Saksela, K., Benichou, S., & Geyer, M. (2014). Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 11(1), 24.

Figure 7 Binding of sdAb19 in Neffin correlates with the inhibition of CD4 internalization.

HeLa-CD4 cells were transfected with plasmids for expression of either Nef-GFP or GFP in combination with the plasmid for expression of wild-type or mutated sdAb19 (1:3 Nef:sdAb19 plasmid ratio). Transfected cells were analyzed for CD4 cell surface expression and cell lysates were analyzed by Western blotting.

Lülf, S., Matz, J., Rouyez, M. C., Järviluoma, A., Saksela, K., Benichou, S., & Geyer, M. (2014). Structural basis for the inhibition of HIV-1 Nef by a high-affinity binding single-domain antibody. Retrovirology, 11(1), 24.