Recombinant Ebola and Influenza Protein Probes

Recombinant Ebola virus glycoprotein with the mucin-like domain deleted (GP_Δmuc), and HIV-1 fusion peptide probe (VRC34-epitope scaffold-FP) and knock-out scaffold probe (VRC34-epitope scaffold-KO) were produced as described previously^{19 ,31 (link),32 (link)}. Proteins were biotinylated and conjugated with streptavidin-APC (GP_Δmuc and VRC34-epitope scaffold-FP) or streptavidin-PE (VRC34-epitope scaffold-KO) (Thermo Fisher Scientific), respectively. Recombinant hemagglutinins (A/California/07/2009, A/Solomon Islands/3/2006, and A/Wisconsin/67/2005) were produced as before^{33 (link)} or acquired from BEI Resources (A/Victoria/210/2009, and B/Brisbane/60/2008) and were biotinylated using an EZ-Link Sulfo-NHS-LC-Biotin kit (Thermo Fisher Scientific). Anti-flag FITC antibody was purchased from Sigma-Aldrich (clone M2).

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Wang B., DeKosky B.J., Timm M.R., Lee J., Normandin E., Misasi J., Kong R., McDaniel J.R., Delidakis G., Leigh K.E., Niezold T., Choi C.W., Viox E.G., Fahad A., Cagigi A., Ploquin A., Leung K., Yang E.S., Kong W.P., Voss W.N., Schmidt A.G., Moody M.A., Ambrozak D.R., Henry A.R., Laboune F., Ledgerwood J.E., Graham B.S., Connors M., Douek D.C., Sullivan N.J., Ellington A.D., Mascola J.R, & Georgiou G. (2018). Functional Interrogation and Mining of Natively-Paired Human VH:VL Antibody Repertoires. Nature biotechnology, 36(2), 152-155.

Publication 2017

streptavidin-APC streptavidin-PE EZ-Link Sulfo-NHS-LC-Biotin kit Anti-flag FITC antibody

Anti antibody Clone Ebola like virus Epitope Fitc Glycoprotein Hemagglutinins Hiv 1 Mucin Peptide Proteins Streptavidin Sulfo nhs lc biotin

Corresponding Organization :

Other organizations : The University of Texas at Austin, National Institute of Allergy and Infectious Diseases, University of Kansas, Harvard University, Boston Children's Hospital, Duke Medical Center

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Variable analysis

independent variables

Recombinant Ebola virus glycoprotein with the mucin-like domain deleted (GP_Δmuc)
HIV-1 fusion peptide probe (VRC34-epitope scaffold-FP)
Knock-out scaffold probe (VRC34-epitope scaffold-KO)
Recombinant hemagglutinins (A/California/07/2009, A/Solomon Islands/3/2006, A/Wisconsin/67/2005, A/Victoria/210/2009, and B/Brisbane/60/2008)

dependent variables

Biotinylation and conjugation of proteins with streptavidin-APC (GP_Δmuc and VRC34-epitope scaffold-FP) or streptavidin-PE (VRC34-epitope scaffold-KO)

control variables

Anti-flag FITC antibody (clone M2) purchased from Sigma-Aldrich

controls

Positive control: None mentioned
Negative control: None mentioned

Annotations

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