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. 2020 May 2;117(7):1990–2007. doi: 10.1002/bit.27355

Figure 5.

Figure 5

ELISA and western blot analysis of recombinant viral antigens. (a) ELISA analysis of IgG titers in mouse serum immunized with mRID‐RBD and mRID‐HR2. mRID‐RBD, mRID‐HR2, mRID (the negative control), and RBD from overexpressing 293T cells (the positive control) were used as coating antigens. (b) ELISA titers of anti‐mRID‐H5gd immune sera. mRID‐H5gd and mRID were used as coating antigens. All mouse sera were twofold serially diluted (MERS‐CoV serum and influenza serum starting from 1:200 and 1:6,400, respectively). All data are presented as mean ± SD (n = 5). (c) RBD‐ and H5gd‐specific polyclonal antibodies were detected by western blot analysis using mRID‐RBD, RBD (from 293T cells; upper panel), and influenza H5gd (lower panel). The target proteins are represented by blue arrows. All proteins were purified using a single Ni–NTA affinity column. (d) Detection of H5N1‐specific antibodies in mouse sera. Egg‐derived HA antigens from H5N1 and H5N8 strains (Green Cross Pharma, Republic of Korea) were used as coating antigens. The sera were serially diluted from 1:100. ELISA, enzyme‐linked immunosorbent assay; IgG, immunoglobulin G; MERS CoV, Middle East respiratory syndrome corona virus; mRID, mouse RNA‐interacting domain; RBD, receptor‐binding domain [Color figure can be viewed at wileyonlinelibrary.com]