AVS 53rd International Symposium
    Biomaterial Interfaces Tuesday Sessions
       Session BI2-TuM

Paper BI2-TuM10
HaloTag@super TM@ Technology for Protein Arrays

Tuesday, November 14, 2006, 11:00 am, Room 2014

Session: Biodiagnostic Innovation
Presenter: N. Nath, Promega Corporation
Authors: N. Nath, Promega Corporation
J. Zhu, Promega Corporation
D. Klaubert, Promega Corporation
D. Storts, Promega Corporation
K. Wood, Promega Corporation
Correspondent: Click to Email
Protein interaction arrays are emerging tools geared toward proteome scale detection of protein-protein, protein-drug or protein-DNA 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 HaloTag@SUPER TM@ technology for rapid and covalent capture of fusion proteins in an oriented fashion directly from complex protein matrices without any prior purification. HaloTag protein is a mutant hydrolase that makes covalent bonds with its chloroalkane substrate. We demonstrate that proteins expressed as HaloTag fusions can be sensitively and irreversibly captured on a PEG based microarray substrate modified with chloroalkane substrate. Because HaloTag is compatible with in-vitro protein expression systems, multiple fusion proteins can be rapidly synthesized (90min) and immobilized in parallel. We also demonstrate that captured fusion proteins are functionally active and are able to interact with their interacting partner. Result indicates that HaloTag technology is perfectly suited for rapid prototyping of protein interaction arrays for large scale study of interaction networks.