Fig. 8 | Scientific Reports

Fig. 8

From: Development of novel canine phage display-derived neutralizing monoclonal antibody fragments against rabies virus from immunized dogs

Fig. 8

SDS‒PAGE and Western blot analysis of the five purified, concentrated, and soluble K9RABVscFv clones. The actual scFv molecular weight was approximately 30–37 kDa (black arrow), and a 60-kDa (white arrow) dimeric form of scFv was observed in four clones. (a) SDS‒PAGE analysis of 5 K9RABVscFv (b) Anti-6XHIS tag-conjugated HRP secondary monoclonal antibody (clone HIS-1, Sigma‒Aldrich, UK) at a concentration of 1:10,000 bound to the scFv antibody in the WB assay and detected by ECL reagents (Cytiva Amersham). (c) All five scFv antibodies incorporated with primary antibody 1:5,000 dilution of unconjugated goat anti-human IgG [F(ab’)2] (Invitrogen; Thermo Fisher Scientific) and a 1:5,000 dilution of HRP-conjugated rabbit anti-goat IgG (H + L) (Invitrogen; Thermo Fisher Scientific) as a secondary antibody. The secondary antibodies were detected via enhanced chemiluminescence (ECL) reagents (Cytiva Amersham). The original Western blot images are shown in Fig. S3. A Spectra Multicolor Broad Range Protein Ladder (Thermo Scientific) was used in panels (a) and (c).

Back to article page