Figure 2 | Scientific Reports

Figure 2

From: A targeted boost-and-sort immunization strategy using Escherichia coli BamA identifies rare growth inhibitory antibodies

Figure 2

Combining native and recombinant BamA formats enables discovery of growth inhibitory pAbs. (a) Schematic of “targeted boost-and-sort” strategy in which Sprague-Dawley (SD) rats are initially primed with K12 bacteria (red arrow) followed by two boosts with the recombinant amphipol-BamA (blue arrow). After subsequent cell fusion to generate hybridomas, fluorescently-labeled BamA is used to single cell sort BamA+ hybridomas to generate a final enriched panel. (b) pAb ELISA titers using purified pAbs dramatically improve post-protein boost (dashed line) compared to pre-boost (solid line). (c) pAbs from three rats (R2, R5, and R6) reduce growth of ΔwaaD E. coli as measured by OD600 density only after protein boost (white vs. black box). Growth differences observed for bleed 2 in R2, R5, and R6 were statistically significant compared to buffer controls (n = 3, p < 0.01).

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