Recombinant Human Anti-RSV Antibody (AM14) (CAT#: PABL-321)

Figure 1 Internalization of Fab fragments.

RSV-infected HEp-2 cells were incubated with RSV F-specific MAbs D25, AM14, 5C4, and MPE8 or corresponding monomeric Fab fragments at the same concentration for 90 min to induce internalization. Afterwards the cells were fixed, permeabilized, and stained with AF488 human anti-goat IgG or AF488 chicken anti-mouse IgG (green). Nuclei were visualized with DAPI (blue). The amount of internalized vesicles was quantified in 50 positive cells.

Leemans, A., De Schryver, M., Van der Gucht, W., Heykers, A., Pintelon, I., Hotard, A. L.,... & Broadbent, L. (2017). Antibody-induced internalization of the human respiratory syncytial virus fusion protein. Journal of virology, 91(14), e00184-17.

Figure 2 Neutralization of laboratory strains and clinical isolates of RSV.

Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.

Figure 3 Binding of AM14 to immobilized RSV F proteins was measured using a Luminex system.

Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.

Figure 4 Binding of AM14 Fab to immobilized prefusion RSV F was measured by surface plasmon resonance. Best fit of the data to a 1:1 binding model is shown in red.

Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.

Figure 5 N426D disrupts binding of AM14 to prefusion F. Relative binding of D25, 101F, MPE8 and AM14 to cell surface-expressed prefusion F (grey) or prefusion F containing the AM14 escape mutation, N426D (white) was measured by flow cytometry. Data were normalized to motavizumab binding. Binding of D25, 101F and MPE8 to N426D was comparable to wild-type prefusion F, whereas AM14 binding to N426D was reduced four-fold.

Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.

Figure 6 AM14 is specific for cleaved, trimeric RSV F.

Binding of antibodies AM14 to uncleaved monomeric RSV F (open black triangles), cleaved monomeric RSV F (black circles), uncleaved postfusion RSV F (open blue triangles), cleaved postfusion RSV F (blue circles), uncleaved prefusion RSV F (open red triangles) and cleaved prefusion RSV F (red circles) was measured by ELISA.

Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.

Figure 7 AM14 stabilizes RSV F trimer in the absence of the foldon trimerization motif. Size-exclusion chromatography profiles from a Superose 6 column are shown for AM14 Fab or D25 Fab complexed with prefusion RSV F containing the foldon trimerization motif (black and grey, respectively) and for AM14 Fab or D25 Fab co-expressed with RSV F ectodomain without foldon (red and blue, respectively).

Gilman, M. S., Moin, S. M., Mas, V., Chen, M., Patel, N. K., Kramer, K.,... & Beaumont, T. (2015). Characterization of a prefusion-specific antibody that recognizes a quaternary, cleavage-dependent epitope on the RSV fusion glycoprotein.PLoS pathogens, 11(7), e1005035.

Figure 1 Recombinant Human Anti-RSV Antibody (PABL-321) in SDS-PAGE

SDS-PAGE analysis of PABL-321 in non-reduced (Lane 2) and reduced (Lane 1) conditions.

Figure 2 Recombinant Human Anti-RSV Antibody (PABL-321) in HPLC

The purity of PABL-321 was greater than 95% as determined by SEC- HPLC.

Figure 3 Recombinant Human Anti-RSV Antibody (PABL-321) in WB

Western blot analysis of PABL-321 was performed by loading Human respiratory syncytial virus (RSV) (A2) Fusion glycoprotein / RSV-F Protein (His Tag) onto a 12% Tris-HCl polyacrylamide gel. Proteins were transferred to a CN membrane and blocked with 5% skim milk for at least one hour. Membranes were probed with PABL-321 and HRP Goat Anti-Human IgG as a secondary antibody. Chemiluminescent detection was performed.
Lane 1: Reduced antigen (1 μg)

Figure 4 Recombinant Human Anti-RSV Antibody (PABL-321) in DB

Dot Blot analysis of PABL-321 was performed by coating with Human respiratory syncytial virus (RSV) (A2) Fusion glycoprotein / RSV-F Protein (His Tag).

PABL-321 incubation concentration: 2 μg/mL.
HRP-conjugated goat anti-Human IgG as a secondary antibody (1: 2000)

Figure 5 Recombinant Human Anti-RSV Antibody (AM14) (PABL-321) in ELISA

ELISA analysis of PABL-321 was performed by coating with Human respiratory syncytial virus (RSV) (A2) Fusion glycoprotein / RSV-F Protein (His Tag).