Preliminary applications of the mAbs
To explore the potential application of these mAbs, three mAbs B2A1,
B2H10, and B2E12, which were the representative of each mAb group, were
used in IFA, immunohistochemical staining, and immunoelectron microscopy
for detecting pB602L in ASFV infected cells or tissues. During the mAb
characterizing process, we proved that all these mAbs were performed
well in detecting recombinant pB602L expressing in Sf-12 cells by IFA.
But when they were used to stain ASFV infected PAM cell, strong
florescence signals were detected only in the cells stained with B2H10
(Fig. 3). In the immunoelectron microscopy test, specific high electron
density gold particles were observed in ASFV assembly factories of the
cells which were labeled with B2A1 (Fig. 4A), B2H10 (Fig. 4B), and B2E12
(Fig. 4C), respectively, but not in the controls labeled with PBS
instead of an antibody (Fig. 4D). Interestingly, we also found that few
gold particles were localized on the surface of viral particles or at
vesicular membranes. In the immunohistochemical test, very strong
positive signs were observed in all the ASFV infected pig tissues
including spleen, tosil, and gastrahepatic lymph node that were stained
with each of the three mAbs, but not in the controls (Fig. 5),
indicating all these mAbs can be used in immunohistochemical staining.