Recombinant Mouse Anti-TGEV Spike protein Antibody (1AF10) (CAT#: PABL-719)

Figure 1 APN-binding domain and epitopes for neutralizing mAbs overlap in TGEV and PRCV S proteins.

A . Scheme of TGEV and PRCV S proteins showing the S1, S2, transmembrane (T) and cytoplasmic (Cy) regions. Location of the C, B, D and A antigenic sites [21], and the pAPN RBD (bar with N and C-terminal residues) [22] are shown. Length is indicated for mature S1 regions. B. A short, soluble S protein variant containing the TGEV RBD region binds to cell surface pAPN. Binding of a bivalent SA-Fc fusion protein (SA) to BHK-pAPN (open histograms), alone (left) or in the presence of the site A-specific 6AC3 mAb (right), as analyzed in FACS. Filled histograms correspond to an unrelated Fc fusion protein. C. Binding of site A-specific TGEV- neutralizing mAb to the SA protein. mAb binding to plastic-bound SA protein, monitored by optical density (OD). Site A mAbs are specific for the Aa (1BB1), Ab (1DE7) and Ac (6AC3, 1AF10) subsites [24]. An anti-HA mAb that binds to the HA tag in the SA protein was used as control. D. Site A-specific mAbs prevent SA protein binding to pAPN. Binding of the SA protein to BHK-pAPN cells in panel B was monitored alone or in the presence of site A (shown in C) or site D-specific (1DG3) mAb, and the binding ratio determined (see Materials and Methods). C, D . Mean and standard deviation for three experiments.

Reguera, J., Santiago, C., Mudgal, G., Ordoño, D., Enjuanes, L., & Casasnovas, J. M. (2012). Structural bases of coronavirus attachment to host aminopeptidase N and its inhibition by neutralizing antibodies. PLoS pathogens, 8(8), e1002859.