Team:Penn/SurfaceDisplayOverview

*Schematic* While engineering bacteria as a light-activated drug delivery system is novel and useful by itself, the targeting potential of our light-activated bacterial therapeutic would be greatly improved if the bacteria also could target specific cells. The goal of the second module of our system is to allow bacterial targeting to cancer cells. We sought to achieve this by displaying an engineered cancer cell binding protein on the surface of our E. coli. We successfully displayed DARPin H10-2-G3, an antibody-mimetic protein rationally evolved to picomolar affinity with HER2, a breast cancer biomarker. We verified that bacteria displaying this protein can selectively bind to breast cancer cells in vitro. This class of protein has not been displayed before on the surface of bacteria. Along the way, we developed a generalized BioBrick surface display system which allows future iGEM teams to display proteins of their choosing on the surface of bacteria.

Objectives

Our goals were twofold:

1. Create a generalized BioBrick surface display platform for other labs and iGEM teams.
2. Use this system to display DARPin H10-2-G3 on the surface of E. coli and verify that the system targets cancer cells.