Research program 4-2
Program area 4, project 2:
The project is focused on development of a new Escherichia coli-based bacterial surface display system for proteins and its use in selection of protein-based affinity reagents from libraries. In the project, display of proteins on the E. coli surface is based on so-called autotransporters, naturally occurring proteins characterized by a capability to direct and present "passenger" proteins to the outside of the cell via an anchoring domain positioned in the outer membrane. Preliminary data support the use of this system for protein library applications, and a number of potential improvements have been identified, which are addressed in the present project to develop the AIDA-I surface display system as a viable option for protein display application of relevance for both the academic and industrial partners involved. These potential improvements include throughput (speed), cell-cell aggregation and optimizations to allow surface display of a wide range of affinity proteins classes of different structure and molecular properties. The main focus during the initial parts of the project will be on the actual selection of binders from the libraries and strategies to implement automated and parallelized magnetic-bead selections (MACS) are investigated.
Project leader: John Löfblom
John Löfblom is an Associate Professor at the Division of Protein Technology at KTH and an expert on protein engineering technology and development of methods for directed evolution. Current research is including engineering of artificial ligands for targeting angiogenesis and tissue engineering, engineering of prodrug formats of affinity proteins and cell therapies, development of methods for directed evolution of protease and protease substrates, engineering of targeting agents for improved uptake into the central nervous system and development of high-throughput selection systems for affinity proteins (part of the CellNova center).
PhD student:
Luke Parks
Project 4-2 CellNova Mars 2021.pdf (pdf 1.4 MB)
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