Polymersomes are stable self-assembled architectures which mimic cell membranes. For characterization, membrane proteins can be incorporated into such bio-mimetic membranes by reconstitution methods, leading to so-called proteopolymersomes. In this work, we demonstrate the direct incorporation of a membrane protein into polymersome membranes by a cell-free expression system. Firstly, we demonstrate pore formation in the preformed polymersome membrane using α-hemolysin. Secondly, we use claudin-2, a protein involved in cell-cell interactions, to demonstrate the in vitro expression of a membrane protein into these polymersomes. Surface plasmon resonance (Biacore) binding studies with the claudin-2 proteopolymersomes and claudin-2 specific antibodies are performed to show the presence of the in vitro expressed protein in polymersome membranes.
19 November 2012 in Angewandte Chemie International Edition
We report on the integration of optical microsensors into a cell culture microchannel device. We demonstrate the possibility of measuring the glucose and oxygen concentrations in the microenvironment of the mammalian cells cultured in a microchannel device. Furthermore, cell proliferation and morphology could be monitored microscopically while these measurements were being made. Through the use of multiple sensors along the length of the microchannel, concentration gradients of various metabolites, such as oxygen, as well as the effects of cell uptake and perfusion rate of growth medium on these gradients could be studied. As such, the system allowed real-time observations of the cells' response to their chemical microenvironment. Our approach allows cell culture and cell assays to be performed simultaneously in an integrated microchannel system with potential applications as a research tool or drug screening method.