Phage display technology for surface functionalization of a synthetic biomaterial.

摘要

The rapid growth in the use of synthetic polymers in medicine and biotechnology has prompted the development of advanced biomaterials that present unique surface properties to control cellular activity. To control the surface properties of biomaterials numerous methods have been developed for immobilization of biomolecules. The goal of this work was to develop a new method for surface functionalization of synthetic biopolymers using phage display technology. This approach has traditionally been utilized for both biological and non-biological materials to select peptides expressed on the bacteriophage using a combinatorial approach. As presented in this thesis chloride-doped polypyrrole (PPyCl) was used as a model biopolymer to screen for a peptide insert selected from a combinatorial library with diversity of 109. A PPyCl-binding peptide (T59) was successfully identified using this phage display approach.; As a biomaterial, polypyrrole presents many unique opportunities in the field of biomedicine, specifically in tissue engineering, drug delivery and biosensor development. A peptide-expressing phage (&phis;T59) that binds to PPyCl, when compared to other selected materials, was identified. Furthermore, the T59 peptide, independent of the phage, was synthesized and its binding ability and characteristics were analyzed using both qualitative and pseudo-quantitative analysis. Furthermore, the stability of the peptides in the presence of serum proteins was explored using indirect methods to compare to a control condition. Finally, we explored a potential application of the selected T59 peptides by attaching a cell-adhesion promoting sequence that permitted cell attachment on PPyCI surface without the presence of serum proteins. Although not directly shown here, this approach, which is highly versatile, simple and imparts not changes to the material's bulk properties, can potentially be applied to various biopolymers.