| AVS 55th International Symposium & Exhibition | |
| Biomaterial Interfaces | Tuesday Sessions |
| Session BI+NC-TuA |
| Session: | Protein and Cells Interactions on Micro- and Nanofabricated Substrates |
| Presenter: | N. Nath, Promega Corporation |
| Authors: | N. Nath, Promega Corporation R. Hurst, Promega Corporation B. Hook, Promega Corporation K. Zhao, Promega Corporation D. Storts, Promega Corporation B. Bulleit, Promega Corporation |
| Correspondent: | Click to Email |
Protein arrays are emerging tools geared toward proteome wide detection of protein-protein, protein-drug, protein-DNA or protein-antibodies interactions. Wide application of protein array technology however faces significant challenge due to lack of high-throughput method for protein expression and purification. Here we present a new integrated approach for creating protein arrays that combines in-vitro protein expression system with HaloTag™ capture technology. The method allows for rapid and covalent capture of HaloTag™ fusion proteins in an oriented fashion directly from complex protein matrices without any prior purification. Multiple fusion proteins can be rapidly synthesized (90min) and immobilized in parallel for high throughput studies. We also demonstrate that arrayed fusion proteins are functionally active and can be used for protein-protein and protein-nucleic acid interaction studies. Furthermore, we show that by using a HaloTag-Protein G fusion we can fabricate antibody arrays directly from ascites fluid without any prior purification of antibodies. Unlike current antibody array platforms, antibodies on our platform are oriented on the surface for maximum biological activity. HaloTag™ protein arrays thus provide a single platform for multiple- biomolecular interaction studies.