Method of fabrication of microarray of gel-immobilized...

Chemistry: molecular biology and microbiology – Apparatus – Including measuring or testing

Reexamination Certificate

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C435S288300, C435S288400

Reexamination Certificate

active

06656725

ABSTRACT:

BACKGROUND OF THE INVENTION
1. Field of the Invention
This invention relates to a method for making polymerized molecules and more particularly, this invention relates to a method for preparing polymer gel grids or arrays on a solid surface via copolymerization wherein each discrete element in the array contains different tethered molecules.
2. Background of the Invention
The manufacturing and use of microchips containing arrays of immobilized oligonucleotides and DNA is a rapidly developing field. Such microchips have been applied for diagnostics of mutations, screening single-nucleotide polymorphisms, identification of microorganisms, monitoring gene expression, as well as for other analytical purposes.
The inventors previously conceptualized and developed microarrays of gel-immobilized compounds on chips (MAGIChips™), containing oligonucleotides, DNA, or proteins. The three-dimensional gel support of MAGIChips provides some essential advantages. The gels have much higher capacity, allowing the immobilized molecules to be well spaced from each other and to interact with the test compound in more homogeneous fashion than when the molecules are attached to solid supports. The glass surface separating the gel pads is treated to become hydrophobic, preventing the exchange of water solution between adjacent gel pads and allowing different chemical and enzymatic reactions to be carried out in the individual gel pads. The gel pads are essentially microtest tubes with volumes ranging from pico-liters to nanoliters.
Oligonucleotide arrays can be fabricated by parallel synthesis directly on a support. See U.S. Pat. No. 5,861,247 awarded to the instant Assignee on Jan. 19, 1999. The photodirected synthesis of oligonucleotides in situ was demonstrated to be highly efficient for manufacturing microchips of high complexity and density.
However, a need exists in the art for a method to produce biochips to accommodate synthetic and natural compounds. The method should utilize standard laboratory solutions in a minimal number of steps to produce the biochips. The method should also minimize any chance for cross contamination or chemical communication between discrete entities on the biochip. A myriad of solid support substrates should also be appropriate.
SUMMARY OF THE INVENTION
It is an object of the present invention to provide a method for polymerizing molecules on a solid substrate that overcomes many of the disadvantages of the prior art.
Another object of the present invention is to provide a simple method for polymerizing monomers into discrete entities. A feature of the invention is that the polymerizing means is dispersed in a vehicle for preventing chemical communication between entities. An advantage of the invention is that the contents of each of entities do not mix with each other.
Still another object of the present invention is to provide a method for making porous polymers containing different molecules tethered therein. A feature of the invention is that the polymers are arranged in close proximity to each other yet contain different molecules, such as oligonucleotides, nucleic acids, and low molecular weight compounds, each of which contain an unsaturated moiety prior to combination with the constituent material comprising the porous polymers. In one protocol, the unsaturated moiety covalently combines with the constituent material in an elimination reaction. Another feature of the invention is that the polymers are produced either via serial application of reactants or a simultaneous application of reactants. An advantage of the invention is that it facilitates rapid production of biochips with a minimal amount of contamination between adjacent polymer structures.
Briefly, the invention provides for a method for making discrete porous polymeric structures wherein each structure contains different molecules, the method comprising contacting a solution containing monomer to a solid substrate so as to form discrete accumulations of the monomer on the substrate; and contacting the accumulations with a means for polymerizing the monomer, wherein the means is dispersed in a vehicle which prevents cross contamination of the accumulations.


REFERENCES:
patent: 5861247 (1999-01-01), Mirzabekov et al.
patent: 5932711 (1999-08-01), Boles et al.
patent: 5981734 (1999-11-01), Mirzabekov et al.
patent: 6174683 (2001-01-01), Hahn et al.
patent: 6183970 (2001-02-01), Okano et al.
patent: 6372813 (2002-04-01), Johnson et al.
Vasiliskov, et al.; “Fabrication of Microarray of Gel-Immobilized Compounds on a Chip by Copolymerization”; _BioTechniques_; Sep. 1999; p. 592-605; vol. 27 No. 3.

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