Recombinant Mouse Anti-RSV gp F Antibody (101F) (CAT#: PABL-694)

Figure 1 Neutralization with anti-F MAbs added before (A) or during (B) HRSV infection.directed against the F protein of HRSV.

(A) Long virus (8.2 × 105 PFU) was incubated in the absence or presence of 400 μg of the anti-F MAbs indicated on the figure. Virus-antibody mixtures were ultracentrifuged (125,812 × g for 2 h in a Beckman SW60 rotor), and the pellets were resuspended and used to infect HEp-2 cells. After 72 h of incubation, production of viral antigen was quantified by ELISA, as described in Materials and Methods, and results are presented as a percentage of the value for control cells infected in the absence of antibody. MAb 1B.C11 against a capsid protein of ASPV was used as a negative control (C−) in neutralization. (B) Long virus (1 × 105 PFU) ultracentrifuged as before in the absence of antibody was used to infect HEp-2 cells in the absence or presence of 3 μg of the antibodies indicated on the figure. Production of viral antigen was quantified as described for panel A. Data represent the mean and standard deviation from three independent experiments.

Magro, M. , Andreu, D. , Paulino GómezPuertas, José A Melero, & Concepción Palomo. (2010). Neutralization of human respiratory syncytial virus infectivity by antibodies and low-molecular-weight compounds targeted against the fusion glycoprotein. Journal of Virology, 84(16), 7970-7982.

Figure 2 Antigen binding and virus neutralization with MAbs directed against the F protein of HRSV.

Serial dilutions of purified anti-F MAbs were tested in a direct ELISA for binding to a soluble form of the F protein (FTM−), as described in Materials and Methods.

Magro, M. , Andreu, D. , Paulino GómezPuertas, José A Melero, & Concepción Palomo. (2010). Neutralization of human respiratory syncytial virus infectivity by antibodies and low-molecular-weight compounds targeted against the fusion glycoprotein. Journal of Virology, 84(16), 7970-7982.

Figure 3 Quantification of virus binding to cells and infectivity in the presence of MAbs.

(A) Long virus (6.5 × 105 PFU) preincubated in the absence or presence of 30 μg of the antibodies indicated on the figure, was mixed with a suspension of HEp-2 cells for 1 h at 4°C, and the virus bound to cells was detected by flow cytometry using an anti-F Cy5 antibody. (B) Aliquots of the virus-antibody-cell mixtures were replated. Forty-eight hours postinfection, cells were resuspended and labeled with an anti-F Cy5 antibody. Mock-infected cells or cells incubated with the MAb 63G against the attachment (G) glycoprotein or with the MAb 1B.C11 (C−) against a capsid protein of ASPV were used as controls in the experiment.

Magro, M. , Andreu, D. , Paulino GómezPuertas, José A Melero, & Concepción Palomo. (2010). Neutralization of human respiratory syncytial virus infectivity by antibodies and low-molecular-weight compounds targeted against the fusion glycoprotein. Journal of Virology, 84(16), 7970-7982.

Figure 4 Inhibition of syncytium formation by anti-F MAbs.

(A) BSR-T7/5 cells were transfected with 500 ng of pTM1 plasmid encoding the full-length F gene. The transfection mixture was removed 7 h later, and the MAbs (40 μg/ml) indicated below each panel were added to the culture. Formation of syncytium was evaluated after incubation for 48 h, as described in Materials and Methods. (B) HEp-2 cells were infected with the Long strain of HRSV (multiplicity of infection, 0.1 PFU/cell). The inoculum was removed at 90 min postinfection, and cells were maintained with medium. Five hours later, MAbs were added to the culture. Syncytium formation was examined as described for panel A. The results shown in this figure are representative of three independent experiments.

Magro, M. , Andreu, D. , Paulino GómezPuertas, José A Melero, & Concepción Palomo. (2010). Neutralization of human respiratory syncytial virus infectivity by antibodies and low-molecular-weight compounds targeted against the fusion glycoprotein. Journal of Virology, 84(16), 7970-7982.