Thesis Transport

Thesis Transport-30
The aim of this thesis was the development of stimulus responsive polymer brush structures as micropore switches for molecular screening.This was achieved by grafting p H-responsive poly(methacrylic acid) (PMAA) brushes from nanoporous platforms that were then integrated in functional assays for membrane proteins.AFM images showed open nanopores at p H 4 and closed ones at p H 8.

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The results of these measurements, combined with diffusion experiments using a fluorescent dye to investigate mechanical gating, demonstrated the controlled transport of ions and small molecules across the pores.The formation and characterization of artificial lipid bilayers and the integration of membrane proteins in the biomembranes is also part of this chapter.Specific examples of the relevance of the polymer support for artificial lipid bilayers are mentioned.The PMAA brush growth and its p H-sensitive behavior on the nanoporous platforms were confirmed by scanning electron microscopy (SEM), atomic force microscopy (AFM) and FTIR spectroscopy.AFM imaging was further used to observe the swelling of the PMAA brush selectively inside the wells upon varying the p H of the liquid environment from 4 to 8.In Chapter 5 the PMAA brush functionalization and characterization of nanoporous platforms with wells (dead-end pores) or channels (pores through) is described.The brushes were synthesized by SI-ATRP in a mixture of water and methanol (1:1 by volume) to improve the wettability of the pores.It was also demonstrated that the protein density could be varied in a wide range without impairing the formation of the lipid bilayer.Although improvements are needed for the electrochemical measurements of membrane protein activity, yet the potential application of the integrated platform for ion channel protein assays is demonstrated.As additional review of the literature background for the work presented in this thesis, an overview on pore-spanning lipid bilayers for functional assays of membrane proteins is presented in Chapter 3.The role of membrane proteins as drug targets is introduced and the current limitations in the development of such assays are discussed.

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