| Popis: |
Functional immobilization and lateral organization of proteins into micro- and nanopatterns is an important prerequisite for miniaturizing analytical and biotechnological devices. In this thesis I present novel and versatile approaches for high contrast surface micropatterning of proteins, artificial membranes and live cells based on maleimide photochemistry. The patterning strategy is carried-out on glass substrates exploiting a poly(ethylene glycol) PEG polymer layer as a compatible scaffold. The flexible PEG cushion prevents unspecific proteins attachment and cell adhesion to surfaces. The versatility of this method is demonstrated by means of different orthogonal chemistries using covalent- and affinity- based interactions for protein immobilization. Furthermore, using maleimide based alkyl-thiol chemistry, I utilized the patterning approach for capturing liposomes and proteoliposomes onto surfaces. Formation of fluid patterned polymer-supported membranes demonstrating lateral diffusion of lipids and proteins was confirmed by biophysical assays. A similar approach was used for micropatterning of transmembrane proteins in surface adhered live cells. |
