Arrays of biological membranes and methods and use thereof

Abstract

The present invention overcomes the problems and disadvantages associated with prior art arrays by providing an array comprising a plurality of biological membrane microspots associated with a surface of a substrate that can be produced, used and stored, not in an aqueous environment, but in an environment exposed to air under ambient or controlled humidities. Preferably, the biological membrane microspots comprise a membrane bound protein. Most preferably, the membrane bound protein is a G-protein coupled receptor, an ion channel, a receptor serine / threonine kinase or a receptor tyrosine kinase.

Description

CROSS-REFERENCE TO RELATED APPLICATION [0001] This application is a Continuation-in-Part of U.S. Ser. No. 09 / 854,786, filed May 14, 2001, the content of which is relied upon and incorporated herein by reference in its entirety, and benefit priority under 35 U.S.C. §120 is hereby claimed.BACKGROUND OF THE INVENTION [0002] DNA microarrays have become an extremely important bioanalytical tool (e.g. for analyzing gene expression); protein microarray technology has, however, lagged behind. The fabrication of protein arrays is challenging because of difficulties associated with preserving the folded conformation of proteins in the immobilized state, and high amounts of non-specific binding to immobilized proteins. As a large fraction of drug targets are membrane bound proteins (e.g., G-protein coupled receptors, ion-channels, etc.), there is an impetus to develop tools for high-throughput screening against membrane bound proteins. Membrane proteins maintain their folded conformation when ...

AI Technical Summary

Benefits of technology

[0012] In certain embodiments, the surface of the substrate is coated. Preferably, the coating is a material that enhances the affinity of the biological membrane microspot for the substrate. A most preferred coating material confers a contact angle ranging from about 15° to 80°.

Problems solved by technology

The fabrication of protein arrays is challenging because of difficulties associated with preserving the folded conformation of proteins in the immobilized state, and high amounts of non-specific binding to immobilized proteins.

Surfaces that bind lipid membranes can be broadly classified into three categories: (i) hydrophobic surfaces (e.g., self-assembled monolayers presenting terminal methyl groups) that support the adsorption of lipid monolayers; these are of limited utility as they cannot be used to incorporate membrane-spanning proteins (Plant, A. L., Langmuir 1999, 15, 5128-5135); (ii) hydrophilic surfaces (e.g., glass surfaces) that bind bilayer-lipid membranes; these are also of limited utility as they can only be used to incorporate membrane-spanning proteins with extra-membrane domains that are less thick than the layer of adsorbed water (˜1 mm) (Groves, J. T. et al., Science 1997, 275, 651-653; and Groves, J. T. et al., Langmuir 1998, 14, 3347-3350); and (iii) amphiphilic surfaces that contain hydrophobic and hydrophilic portions and bind bilayer-lipid membranes; these offer the potential for incorporating a wide variety of membrane-spanning proteins (Lang, H. et al., Langmuir 1994, 10, 197-210; Raguse, B. et al, Langmuir 1998, 14, 648-659; and Vanderah, D. J. et al, Materials Research Society Fall Meeting Abstracts, Boston, 1999).

However, these methods are cumbersome and require the fabrication of patterned surfaces.

The methods used by Boxer et al., however, have certain limitations.

Second, lipids adsorbed on the bare-glass substrates used by Boxer and co-workers spontaneously desorbed when drawn through an air-water interface (Cremer 1999).

Such systems may not be practical for robust, high throughput, microarray based assays.

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