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.

A fortéBio Octet Red384 instrument was used to measure sera recognition of DS-Cav1, DS-Cav-1 antigenic site Ø KO [15] or DS-Cav-1 antigenic site II KO [15] probes. All assays were performed at 30°C in tilted black 384-well plates (Greiner Bio-One, NC) with agitation set to 1,000 rpm in PBS supplemented with 1% BSA and a well volume of 55 μl. DS-Cav1 and DS-Cav1 KO probes (50 μg/ml) were immobilized for 300 s on HIS1K biosensor tips and equilibrated for 60 s in PBS + 1% BSA prior to measuring association with sera diluted 1:200 in PBS supplemented with 1% BSA. The association after 300 s was recorded.