Chemically modified biological molecules and methods for coupling biological molecules to solid support

Abstract

The invention relates to novel chemically modified biological molecules with enhanced lability towards solid supports, such as glass. These modified molecules can be readily affixed to solid supports, for instance, a glass surface, without first derivatizing the glass surface. High-density microarrays based on these modified molecules as well as methods for preparing these microarrays are also useful.

Description

FIELD OF THE INVENTION The present invention claims a closely related family of compounds, devices, and methods relating to techniques for immobilizing biological molecules to a solid support for the purpose of conducting scientific investigation or routine testing upon the bound molecule samples in areas such as genome-wide genetic mapping and gene expression studies, protein interaction studies, peptide interaction studies & small molecule interactions with larger macromolecules. BACKGROUND OF THE INVENTION A large percentage of investigation in the biochemical arts are directed to studies involving nucleic acids, particularly deoxyribonucleic acid, or DNA. DNA is a water-soluble compound, that if left in solution (i.e., a water-based solution), is likely to degrade, through hydrolysis, and so forth. Obviously this frustrates any investigation involving DNA, and so therefore, accurate and reliable study involving DNA requires a method or device to ensure the integrity of DNA. To...

AI Technical Summary

Benefits of technology

The present invention possesses numerous advantages over the prior art. Many of the advantages derive from the fact that the solid surface, which is typically ordinary glass, remains highly chemically inert. Thus the previously mentioned problems of probe (or other reactant) sticking to the glass .eliminated. The ultimate result is, among other things, far higher detection sensitivity compared with state-of-the-art derivatized solid support due to the absence of non-specific probe absorption.

In addition, the biological molecule to be immobilized upon the solid support is readily derivatized since the reaction conditions can take place in liquid phase for as long as necessary to obtain the required level of derivatization. The reaction of the epoxide derivatives of the present invention is simply to execute it under mild conditions, reaction rates are quick, and equilibrium is highly favorable. Moreover, the epoxide-modified molecule of the present invention is essentially permanently stable, thus it can be prepared and stored for later use. Additional, more specific advantages will be disclosed later during discussion of particular embodiments of the present invention.

Problems solved by technology

Obviously this frustrates any investigation involving DNA, and so therefore, accurate and reliable study involving DNA requires a method or device to ensure the integrity of DNA.

Perhaps the major problem associated with immobilizing DNA on a solid support is exactly how to do it without altering the DNA (other than that relatively small portion that is actually bound to the solid support).

This is a very difficult problem because whatever solid support is used must be essentially inert.

Unfortunately, derivatizing the otherwise inert surface of glass creates problems which could confound the results of the laboratory study involving DNA.

One problem is that derivatization activates and sometime creates a net positive electrostatic charge on the glass surface.

Since DNA is (net) negatively charged, other DNA (or DNA used later in the study but not deliberately affixed to the glass surface) is prone to stick (by non-specific electrostatic attraction) to the glass surface.

Yet simple experiments of this type (probe studies) are severely confounded by electrostatic sticking of the probe to the derivatized (hence electrostatically charged) glass surface.

Yet the radiation detector is unable to distinguish between probe that is chemically bound to a complementary strand of DNA affixed to the solid support, and probe that is simply electrostatically stuck to the glass surface (but not to a DNA strand).

Second, derivatized surfaces result in what shall be known as “spreading.” Spreading occurs because the solid support surface becomes hydrophilic upon derivatization.

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