Recombinant Mouse Anti-HPV16 L2 Antibody (14H6) (CAT#: HPAB-AP368-YC)

Figure 1 Screening for L2 neutralizing monoclonal antibodies (mAbs) using HPV16L2 C50 antigen.

Neutralization geometric mean titers (GMTs) of mAbs 14H6, 15H5, 15E4, 6H8 mAbs were plotted on the log-scaled y-axis against various genotypes of HPV-PsVs on the x-axis. The bar graph was prepared using Prism GraphPad 5.0. All mAb samples were adjusted to an initial concentration of 1.0 mg/ml prior to two-fold serial dilutions. Flow cytometry was used to detect the number of HPV-pseudovirus-infected enhanced green fluorescent protein (EGFP)-expressing cells.

Wang, D., Li, Z., Xiao, J., Wang, J., Zhang, L., Liu, Y., ... & Yu, H. (2015). Identification of broad-genotype HPV L2 neutralization site for pan-HPV vaccine development by a cross-neutralizing antibody. PLoS One, 10(4).

Figure 2 Characterization of 14H6a–f chimeras.

EC50 test by ELISA using a serial dilution of mAb 14H6. The reaction curves vs. the concentration of mAb 14H6 were fitted to a sigmoid trend and the EC50 values were calculated in terms of the half points (shown in the brackets). Detection in western blotting and the EC50 results indicate that all chimeric proteins present mAb 14H6 with similar binding capabilities.

Wang, D., Li, Z., Xiao, J., Wang, J., Zhang, L., Liu, Y., ... & Yu, H. (2015). Identification of broad-genotype HPV L2 neutralization site for pan-HPV vaccine development by a cross-neutralizing antibody. PLoS One, 10(4).

Figure 3 Immunoelectron microscopy of recombinant bacteria.

EM images of ER0808 cells, ER0808-prL8L2 cells, ER2566-pF-L2 cells and ER2566-pC-L2 cells as indicated by gold-conjugated 14H6 (set b, top panel) at ×49,000 magnification. For clarity, close-up views of the top panels are shown; red boxes indicate the magnified regions. The curve of some images below is adjusted to better visualize the gold particles by Photoshop software.

Chen, T., Wang, K., Chi, X., Zhou, L., Li, J., Liu, L., ... & Zhang, J. (2019). Construction of a bacterial surface display system based on outer membrane protein F. Microbial cell factories, 18(1), 70.

Figure 4 Identifcation of fusion protein expression and purifcation.

SDS-PAGE and western blotting of total proteins (TP) and OMPs prepared from equal biomass of ER0808 and a ER0808-prL8HB, ER2566(pF-HB) and ER2566(pC-HB) or b ER0808-prL8L2, ER2566(pF-L2) and ER2566(pC-L2). Black arrows indicate the fusion proteins. Samples (10 µL) with an equivalent amounts of cell lysate, were loaded.

Chen, T., Wang, K., Chi, X., Zhou, L., Li, J., Liu, L., ... & Zhang, J. (2019). Construction of a bacterial surface display system based on outer membrane protein F. Microbial cell factories, 18(1), 70.

Figure 5 Flow cytometry of recombinant bacteria.

E6F6 epitope display strains (a) and E6F6 epitope display strains (b) were analyzed with E6F6 or 14H6 as the primary antibodies and FITC-conjugated secondary antibodies in fow cytometer. ER2566 and ER0808 served as negative control. The results were illustrated as histogram mode and positive cell ratios were indicated.

Chen, T., Wang, K., Chi, X., Zhou, L., Li, J., Liu, L., ... & Zhang, J. (2019). Construction of a bacterial surface display system based on outer membrane protein F. Microbial cell factories, 18(1), 70.