Figure 5.

Binding of HIV-1 gp41 HR2 Antibodies to MAPK14

(A) Representative ELISA graphs show the reactivity of blood anti-gp41 antibodies against overlapping peptides covering the extracellular portion of gp41 (gp41^EC). The sequences of the peptides recognized are indicated on the top.

(B) Representative ELISA graphs show the binding of biotinylated mucosal anti-gp41 to YU-2 gp140 in the presence of potential competitor antibodies (blood-derived anti-gp41 IgG). Error bars indicate the SEM of duplicate values.

(C) Protein microarray plots show the reactivity profile of blood anti-gp140 antibodies against human proteins. For each protein spot, the mean fluorescence intensity (MFI) given by the reference (mGO53) and test antibody are depicted on the y and x axes, respectively. Each dot represents the average of duplicate array proteins. The diagonal lines indicate equal binding for reference and test antibodies.

(D) Bar graphs comparing the MFI values (Left) and Z scores (Right) for the binding of the selected anti-gp41 antibodies to MAPK14 immobilized on the protein array.

(E) Representative ELISA graph comparing the reactivity of blood anti-gp140 antibodies to purified MAPK14 proteins. The 6-195 and mGO53 (green) were used as positive and negative control, respectively. Error bars indicate the SEM of duplicate values.

(F) Affinity of anti-gp41 antibodies to MAPK14. (Left) Representative SPR curves comparing the binding overtime of selected anti-gp41 IgGs (at 250 nM) to purified MAPK14 immobilized on the sensor chip (250 RUs). (Right) Dot plots comparing the relative affinity (K_D) of anti-gp41 antibodies to MAPK14. No binding could be detected (ND) for the following antibodies: 5-25, 10-137, 10-437, and 10-1304.

See also Figure S6.