Generation of Anti-Boa Immunoglobulin Antibodies for Serodiagnostic Applications, and Their Use to Detect Anti-Reptarenavirus Antibodies in Boa Constrictor
Fig 5
Screening of sera from Boa constrictors with BIBD for antibodies against reptarenaviruses by immunoblotting.
Protein pools of Ni-NTA affinity purified recombinant UHV NP (the full-length protein, and N- and C-terminal fragments) separated by SDS-PAGE under reducing conditions were used as the antigens. The membranes were probed with snake serum, and detection of binding was done using either anti-IgY or anti-IgM in combination with an IRDye800 labelled secondary antibody. The immunoblot signals were recorded with Odyssey infrared imaging system (LI-COR Biosciences). The snake serum (or blood) samples #1–4 to #6–9 correspond to the animal numbers of our earlier study (see Table 1, and [20]). The left lanes show the molecular weight marker (Precision Plus Protein Dual Color, Bio-Rad). The arrows indicate the size of the recombinant proteins.