Selection of Single-Domain Antibodies towards Western Equine Encephalitis Virus

Figures

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  Figure A1. Representative gel-filtration chromatography trace.

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  Figure A2. Representative surface plasmon resonance data.

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  Figure 1. Deduced protein sequences of the seven single-domain antibodies (sdAb) that were evaluated. Sequences are given in single letter amino acid code. Alignment was performed using Multalin; high homology positions are shown in red, where lower homology is in blue [48].

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  Figure 2. MagPlex direct binding assay to evaluate the specificity of the putative anti-WEEV E3E2 binding sdAb. Each sdAb was biotin labeled and mixed with MagPlex microspheres coated with various antigens at a range of concentrations. Biotinylated sdAb are denoted by Bt-clone name. Median fluorescence intensity versus Bt-sdAb concentration is plotted. The anti CHIKV sdAb CC3 was also tested as a positive control for the activity of the other sets of microspheres [49].

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  Figure 3. MagPlex sandwich fluoroimmunoassay to evaluate the utility of the sdAb to function as both a capture and recognition reagent for the detection of WEEV E3E2 protein. Only the four best Bt-sdAb recognition reagents are shown. Median fluorescence intensity versus WEEV E3E2 concentration is plotted; error bars represent the SEM.