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Figure 4 | Microbial Cell Factories

Figure 4

From: An engineered autotransporter-based surface expression vector enables efficient display of Affibody molecules on OmpT-negative E. coli as well as protease-mediated secretion in OmpT-positive strains

Figure 4

Evaluation of OmpT-mediated release and small-scale purification of displayed Affibody molecule. A. Flow-cytometric analysis for comparison of surface expression of recombinant proteins between pAraBAD-ZIgG-EC in OmpT-negative E. coli BL21 and pAraBAD-ZIgG-EC in OmpT-positive E. coli DH5α. B. SDS-PAGE showing IMAC-purified supernatants from pAraBAD-ZIgG-EC in OmpT-negative E. coli BL21 and pAraBAD-ZIgG-EC in OmpT-positive E. coli DH5α. Supernatants from non-induced samples, indicated with (−), were included as controls. C. Mass spectrum from ESI-MS analysis of IMAC-purified supernatants from pAraBAD-ZIgG-EC in OmpT-positive E. coli DH5α. Theoretical molecular weight of the OmpT-cleaved ZIgG-His6 is 9712 Da. D. SPR-based biosensor analysis on the IMAC-purified ZIgG-His6. Response units (RU) on the y-axis and time on the x-axis. Representative sensorgrams from injection of ZIgG-His6 at three different concentrations over human IgG immobilized on the chip surface. Injections were performed in duplicates.

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