Structure-Based Design of Head-Only Fusion Glycoprotein Immunogens for Respiratory Syncytial Virus

Boyington, Jeffrey C.; Joyce, M. Gordon; Sastry, Mallika; Stewart-Jones, Guillaume B. E.; Chen, Man; Kong, Wing-Pui; Ngwuta, Joan O.; Thomas, Paul V.; Tsybovsky, Yaroslav; Yang, Yongping; Zhang, Baoshan; Chen, Lei; Druz, Aliaksandr; Georgiev, Ivelin S.; Ko, Kiyoon; Zhou, Tongqing; Mascola, John R.; Graham, Barney S.; Kwong, Peter D.

For the competition neutralization assay, 6 μg of probe (either DS-Cav-1, DS-Cav-1 antigenic site Ø knock out (KO) [15] or DS-Cav-1 antigenic site II KO [15] ) were added to sera and incubated at 37°C for one hour followed by the addition of an equal volume of virus and incubation for an additional hour at 37°C. 50 μl of the sera-RSV F-RSV A2 mixture were added to 2.4 x10 4 HEp-2 cells per well in a 384-well assay plate, and incubated at 37°C for 22-26 hours. The fluorescence intensity for each well was read as described above. One reading was obtained for each mouse sera sample.