SARS-CoV-2 RBD and NTD Biotinylation

Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD and NTD were biotinylated using the Biotin-Protein Ligase-BIRA kit according to the manufacturer’s instructions (Avidity) as described before (Robbiani et al., 2020 (link)). Ovalbumin (A5503-1G; Sigma-Aldrich) was biotinylated using the EZ-Link Sulfo-NHS-LC-Biotinylation kit according to the manufacturer’s instructions (Thermo Fisher Scientific). Biotinylated ovalbumin was conjugated to streptavidin-BB515 (564453; BD). RBD was conjugated to streptavidin-PE (554061; BD Biosciences) and streptavidin-AF647 (405237; Biolegend; Robbiani et al., 2020 (link)). NTD was conjugated to streptavidin-BV421 (405225; Biolegend) and streptavidin-BV711 (563262; BD Biosciences).

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Wang Z., Muecksch F., Muenn F., Cho A., Zong S., Raspe R., Ramos V., Johnson B., Ben Tanfous T., DaSilva J., Bednarski E., Guzman-Cardozo C., Turroja M., Millard K.G., Tober-Lau P., Hillus D., Yao K.H., Shimeliovich I., Dizon J., Kaczynska A., Jankovic M., Gazumyan A., Oliveira T.Y., Caskey M., Bieniasz P.D., Hatziioannou T., Kurth F., Sander L.E., Nussenzweig M.C, & Gaebler C. (2022). Humoral immunity to SARS-CoV-2 elicited by combination COVID-19 vaccination regimens. The Journal of Experimental Medicine, 219(10), e20220826.

Publication 2022

Biotin-Protein Ligase-BIRA kit EZ-Link Sulfo-NHS-LC-Biotinylation kit streptavidin-BB515 streptavidin-PE streptavidin-AF647 streptavidin-BV421 streptavidin-BV711

Af647 Biotin ligase Biotinylation Ovalbumin Protein Sars cov 2 Streptavidin Sulfo nhs

Corresponding Organization : Howard Hughes Medical Institute

Other organizations : Berlin Institute of Health at Charité - Universitätsmedizin Berlin

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Variable analysis

independent variables

Biotinylation of SARS-CoV-2 Wuhan-Hu-1 RBD and NTD using the Biotin-Protein Ligase-BIRA kit
Biotinylation of ovalbumin using the EZ-Link Sulfo-NHS-LC-Biotinylation kit
Conjugation of biotinylated ovalbumin to streptavidin-BB515
Conjugation of RBD to streptavidin-PE and streptavidin-AF647
Conjugation of NTD to streptavidin-BV421 and streptavidin-BV711

dependent variables

Not explicitly mentioned

control variables

Purified and Avi-tagged SARS-CoV-2 Wuhan-Hu-1 RBD and NTD

controls

Ovalbumin (A5503-1G; Sigma-Aldrich) as a control for biotinylation and conjugation procedures

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