Accordingly, MAb 2C7 IgG exhibited bactericidal activity against all 62 minimally passaged gonococcal isolates tested from Nanjing, China (28)

Accordingly, MAb 2C7 IgG exhibited bactericidal activity against all 62 minimally passaged gonococcal isolates tested from Nanjing, China (28). ? 2021 Parzych et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S2. Bactericidal activity of 2C7 DMAb variants expressed in Jh mice. (A) functionality of 2C7-DMAb sera (Jh mice; 3 or 4/group). strain FA1090 was HJC0350 incubated with D3 (left) or D21 (right) sera of mice given DMAb at a final concentration of 33% (vol/vol) (circles/open bars) or 67% (vol/vol) (squares/hatched bars) in the presence of human complement (16.7% [vol/vol]). The graph HJC0350 FGS1 depicts the percent bacterial survival afforded by each serum sample as well as the group averages ( standard HJC0350 deviation [SD]) relative to control plates. (B) Correlation of anti-LOS titers versus bactericidal activity. Anti-LOS titers in the same D3 (left) and D21 (right) serum samples were determined via ELISA using plates coated with purified LOS antigen (strain 15253). Bound antibodies were detected with anti-human IgG-AP and quantified against a standard curve using purified recombinant 2C7 MAb. The graph depicts the 2C7 concentrations (axis) and bactericidal activity (axis) for individual serum samples, which are colored according to DMAb group. These parameters were analyzed using a Spearman correlation (strain 15253. HJC0350 Bound antibodies were detected with anti-human IgG-AP and quantified against a standard curve using purified recombinant 2C7 MAb. Graphs depict the individual 2C7 DMAb concentrations found in washes; the horizontal bars represent group averages. Download FIG?S3, TIF file, 0.09 MB. Copyright ? 2021 Parzych et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S4. Designs of 2C7 DMAb efficacy study using the vaginal colonization model in JHD mice. (A) DMAb administration. Groups of JHD mice (9 or 10/group) were given the indicated 2C7 DMAb plasmids (or pVax control plasmid). All mice were bled throughout the experiment (red drops) to monitor 2C7 expression kinetics (Fig.?4). (B) Primary challenge. On D8 post-DMAb administration, mice were challenged with (FA1090 strain; 4.2??107 CFU, intravaginally), and infection burden was monitored daily (enumeration of vaginal CFU). Bacterial control (daily CFU) and time to clearance were calculated for each treatment HJC0350 group (Fig.?3). (C) Secondary challenge (rechallenge). On D65 postadministration, mice that had had undergone primary challenge and were in the diestrus phase of the estrus cycle and therefore could be infected with (5/group) were rechallenged with (FA1090 strain; 3.5??107 CFU, intravaginally). Infection was monitored as described above. Download FIG?S4, TIF file, 0.2 MB. Copyright ? 2021 Parzych et al. This content is distributed under the terms of the Creative Commons Attribution 4.0 International license. FIG?S5. Serum DMAb expression levels over time in dose response of chimeric 2C7_E430G recombinant IgG (rIgG). Purified 2C7_E430G IgG was produced in CHO cells using traditional bioprocessing strategies and administered systemically at the indicated dose (1, 5, or 25 g/animal; (D1) and infection was monitored via daily vaginal swabbing (Fig.?5A). (A) Bacterial burden following challenge. Graphs depict the average log10 CFU (mean SEM) detected in the vaginal mucosa on the indicated days postinfection. (B) Time to bacterial clearance is shown using Kaplan-Meier curves, which display the percentage of each group with detectable vaginal CFU on the given day post-challenge. Pairwise comparisons were made with the Mantel-Cox log-rank test, and significance was set at 0.008 (Bonferronis correction for 4 groups). The following comparisons were significant: saline versus 2C7_E430G (1 g), = 0.002. (C) Overall bacterial control in each animal was assessed using AUC (log10 CFU) analysis. AUC values for each.