Seminars Archive
Surface Display – An Easy Method for Difficult Tasks
Institute for Pharmaceutical and Medicinal Chemistry, Westfälische Wilhelms-Universität Münster, Germany
Abstract
Displaying a protein of interest on the surface of a living cell bears several advantages. In the case of enzymes, neither the substrate nor the product of the enzymatic reaction needs to cross a membrane barrier. The enzyme being linked to the cell can be separated from the reaction mixture and hence the product by simple centrifugation. Furthermore, surface display of active enzymes or dimeric proteins facilitates screening of inhibitors without the need of prior purification of the target proteins. Moreover, the surface display of recombinant human Hyaluronidase-1 (Hyal-1) on Gram negative bacteria leads to an active enzyme, while the purification of Hyal-1 out of prokaryotic bacteria leads to inclusion bodies and thus to an inactive enzyme.
Taking these advantages into account, displaying various proteins, such as full length antibodies, kinases or small peptides on the surface of Gram negative bacteria can be a powerful tool to overcome different hurdles and enables the measurement of enzymatic activities and the high throughput screening for inhibitors.