Methodology article

Cell membrane array fabrication and assay technology

Victoria Yamazaki¹ , Oksana Sirenko¹ , Robert J Schafer¹ , Luat Nguyen¹ , Thomas Gutsmann² , Lore Brade² and Jay T Groves^1,3

¹  Synamem Corporation, 863 Mitten Road – Suite 101, Burlingame, CA94010, USA

²  Division of Medical and Biochemical Microbiology, Research Center Borstel, Center for Medicine and Biosciences, Borstel, Germany

³  Department of Chemistry, 109 Lewis Hall, University of California – Berkeley, Berkeley, CA 94720, USA

author email corresponding author email

BMC Biotechnology 2005, 5:18doi:10.1186/1472-6750-5-18

Published:	16 June 2005

Abstract

Background

Microarray technology has been used extensively over the past 10 years for assessing gene expression, and has facilitated precise genetic profiling of everything from tumors to small molecule drugs. By contrast, arraying cell membranes in a manner which preserves their ability to mediate biochemical processes has been considerably more difficult.

Results

In this article, we describe a novel technology for generating cell membrane microarrays for performing high throughput biology. Our robotically-arrayed supported membranes are physiologically fluid, a critical property which differentiates this technology from other previous membrane systems and makes it useful for studying cellular processes on an industrialized scale. Membrane array elements consist of a solid substrate, above which resides a fluid supported lipid bilayer containing biologically-active molecules of interest. Incorporation of transmembrane proteins into the arrayed membranes enables the study of ligand/receptor binding, as well as interactions with live intact cells. The fluidity of these molecules in the planar lipid bilayer facilitates dimerization and other higher order interactions necessary for biological signaling events. In order to demonstrate the utility of our fluid membrane array technology to ligand/receptor studies, we investigated the multivalent binding of the cholera toxin B-subunit (CTB) to the membrane ganglioside GM₁. We have also displayed a number of bona fide drug targets, including bacterial endotoxin (also referred to as lipopolysaccharide (LPS)) and membrane proteins important in T cell activation.

Conclusion

We have demonstrated the applicability of our fluid cell membrane array technology to both academic research applications and industrial drug discovery. Our technology facilitates the study of ligand/receptor interactions and cell-cell signaling, providing rich qualitative and quantitative information.