Human Anti-K. pneumoniae MrkA Recombinant Antibody (clone 1) (CAT#: FAMAB-0609WJ)

Figure 1 Anti-MrkA antibody binding is influenced by the antigen presentation format.

MrkA protein coated directly to the ELISA plate was recognized differently by the anti-MrkA antibodies.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 2 Anti-MrkA antibody binding is influenced by the antigen presentation format.

MrkA protein captured by streptavidin after biotinylation was recognized differently by the anti-MrkA antibodies.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 3 Western blot against full length MrkA and deletion mutants.

Protein samples were resolved on a 4-12% SDS-PAGE gel under reducing condition and subjected to Western blot analysis by a murine anti-his mAb, KP3, and clones 1-6 IgGs as indicated underneath each blot. Sample arrangement is as follow: Lane 1, cell lysate from KP strain 43816DM; Lane 2, E.coli BL21 strain control; Lane 3-5 BL21 strain expressing his-tagged recombinant MrkA, MrkA with N terminal 40 amino acid deletion, and MrkA with C terminal 32 amino acid deletion, respectively. In the anti-his blot, a bracket indicates the positions of the monomeric, full length MrkA as well as the deletion mutants.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 4 Epitope analysis by BLI.

Epitope binning was performed against three test article including KP3, clone 4 and 5 as described in the materials and method. There are three phases (I, II, and III) for each graph. In phase I biotin-labeled MrkA protein was loaded to the neutravidin probe. In phase II one of the mAbs (test articles) as indicated above each graph bound to the immobilized MrkA. In phase III different mAbs as indicated were mixed with the test article mAb and loaded to the probe.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 5 Serotype-independent binding to KP strains by the new antibodies.

Flow cytometry experiment was used to gauge the binding of the four new antibodies against three WT KP strains of different serotype as indicated. KP3 serves as the positive control and R347 is a human IgG1 isotype control.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 6 Serotype independent OPK activity by the new antibodies.

KP strain with LPS serotype O1 was used in the OPK assay. All newly isolated antibodies displayed comparable OPK activities to that of KP3. R347 is a human IgG1 isotype control.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 7 Serotype independent OPK activity by the new antibodies.

KP strain with LPS serotype O2 was used in the OPK assay. All newly isolated antibodies displayed comparable OPK activities to that of KP3. R347 is a human IgG1 isotype control.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 8 In vivo protection activity.

The newly identified antibodies were tested in in vivo challenge models under both prophylaxis and therapeutic settings. In the prophylaxis setting antibodies were given 24 hours prior to KP challenge. There were eight animals in each group. R347 is a human IgG1 isotype control.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 9 In vivo protection activity.

The newly identified antibodies were tested in in vivo challenge models under both prophylaxis and therapeutic settings. In the therapeutic setting antibodies were given one hour after KP challenge. There were eight animals in each group. R347 is a human IgG1 isotype control.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura

Figure 10 No synergistic effect by antibody combinations in the therapeutic model.

KP3 was combined with either clone 1 or 5 in equal amount as indicated and tested in a therapeutic model. No significant improvement was observed. There were eight animals in each group. R347 is a human IgG1 isotype control.

Wang, Q., Chen, Y., Cvitkovic, R., Pennini, M. E., Chang, C. S., Pelletier, M., ... & Xiao, X. (2017). Anti-MrkA monoclonal antibodies reveal distinct structura