| Method for attachment of silylated molecules to glass surfaces -> Monitor Keywords |
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Method for attachment of silylated molecules to glass surfacesRelated Patent Categories: Chemistry: Molecular Biology And Microbiology, Measuring Or Testing Process Involving Enzymes Or Micro-organisms; Composition Or Test Strip Therefore; Processes Of Forming Such Composition Or Test Strip, Involving Nucleic AcidMethod for attachment of silylated molecules to glass surfaces description/claimsThe Patent Description & Claims data below is from USPTO Patent Application 20070172872, Method for attachment of silylated molecules to glass surfaces. Brief Patent Description - Full Patent Description - Patent Application Claims
CROSS-REFERENCE [0001] This application claims the benefit of U.S. Provisional application No. 60/383,564, filed May 28, 2003, which is incorporated by reference in its entirety. BACKGROUND OF THE INVENTION [0002] Surface modification plays an important role in micro-array biomolecule detection technology for controlling backgrounds and spot morphology. Several modifications were developed using different type of commercially available silanes such as silyl amines, aldehydes, thiols etc. for immobilization of biomolecules such as oligonucleotides. After coating the surface with reactive silanes, the next challenge is immobilization of required biomolecules on the modified surface. The surface loadings always vary with different silanes and even same silane may not give reproducible results. Reproducibility of optimum surface loading has always been a great challenge in this field since surface loading dictates the performance of the assay. Even with simple linear molecules for immobilization, the optimum loading on the surface is difficult to achieve. [0003] Attaching DNA to a modified glass surface is a central step for many applications in DNA diagnostics industry including gene expression analysis. In general, DNA can be attached to a glass surface either through non-covalent, ionic interactions, or through multi-step processes or simple coupling reactions. Several methods have been reported in the literature using glass surface modified with different types of silylating agents.sup.1-6. All these reported methods involve silylating step which uses expensive reagents and analytical tools. Also, these methods are also multi-step processes that are labor intensive and expensive.sup.8-9. Earlier reported methods have involved a laborious synthesis and time consuming procedure.sup.7. Indeed, many of the current immobilization methods suffer from one or more of a number of disadvantages. Some of these are, complex and expensive reaction schemes with low oligonucleotide loading yields, reactive unstable intermediates prone to side reactions and unfavorable hybridization kinetics of the immobilized oligonucleotide. The efficient immobilization of oligonucleotides or other molecules on glass surface in arrays requires a) simple reliable reactions giving reproducible loading for different batches, b) stable reaction intermediates, c) arrays with high loading and fast hybridization rates, d) high temperature stability, e) low cost, f) specific attachment at either the 5'- or 3'-end or at an internal nucleotide and g) low background. [0004] The present invention represents a significant step in the direction of meeting or approaching several of these objectives. SUMMARY OF THE INVENTION [0005] The present invention fulfills the need in the art for methods for the attachment of molecules such as oligonucleotides onto unmodified surfaces such as a glass surface without the need for laborious synthetic steps, with increase surface loading densities, and with greater reproducibility and which avoids the need for pre-surface modifications. Molecules such as DNA can be silylated at either the 3' or 5' ends as discussed below and the 3' or 5'-silylated DNA may then be covalently attached directly to a surface such as a pre-cleaned glass surface (Scheme) for use in hybridization assays. Furthermore, thorough the use of certain silylating reagents, it is now possible to further enhance surface loading densities by using modified silylating agents having multiple molecules attached thereto. The present invention thus provides novel methods for attaching molecules onto a substrate, devices prepared by such methods, and compositions. This method provides great advantages over the present technology in terms of simplicity, cost, speed, safety, and reproducibility. [0006] Thus, in one embodiment of the invention, a method is provided for immobilizing a molecule onto a surface, said method comprising the steps of: [0007] (a) contacting the molecule with an agent so as to form a reactive intermediate, said agent having a formula i: (R.sub.1)(R.sub.2)(R.sub.3)Si--X--NCY i wherein R.sub.1, R.sub.2 and R.sub.3 independently represents C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; X represents linear or branched C.sub.1-C.sub.20 alkyl or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; and Y represents oxygen or sulfur, with the proviso that at least one of R.sub.1, R.sub.2 or R.sub.3 represents C.sub.1-C.sub.6 alkoxy; and [0008] (b) contacting the reactive intermediate with said surface so as to immobilize the molecule onto said surface. [0009] In one aspect of this embodiment, a method is provided for immobilizing a molecule onto a glass surface. [0010] In another embodiment of the invention, a method is provided for immobilizing a molecule onto a surface, said method comprising the steps of: [0011] (a) contacting Si(NCY).sub.4 wherein Y represents oxygen or sulfur with an agent so as to form a first reactive intermediate, said agent having a formula ii: (R.sub.1)(R.sub.2)(R.sub.3)Si--X-Z ii wherein R.sub.1, R.sub.2 and R.sub.3 independently represents C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; X represents linear or branched C.sub.1-C.sub.20 alkyl or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; and Z represents a hydroxy or amino group, with the proviso that at least one of R.sub.1, R.sub.2 or R.sub.3 represents C.sub.1-C.sub.6 alkoxy; [0012] (b) contacting the first reactive intermediate with a molecule so as to form a second reactive intermediate; [0013] (c) contacting the second reactive intermediate with said surface so as to immobilized the molecule onto said surface. The method allows for the production of branched captured molecules structures such as branched oligonucleotides on a surface which is useful for enhancing detection of target analytes such as nucleic acids. [0014] In one aspect of this embodiment of the invention, a method is provided for immobilizing a molecule onto a glass surface. [0015] In another embodiment of the invention, a compound is provided having the formula iii: (R.sub.1)(R.sub.2)(R.sub.3)Si--X--NHCYL-M iii wherein R.sub.1, R.sub.2 and R.sub.3 independently represents C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; X represents linear or branched C.sub.1-C.sub.20 alkyl or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; L represents a linking group; and M represents a molecule, with the proviso that at least one of R.sub.1, R.sub.2, or R.sub.3 represent C.sub.1-C.sub.6 alkoxy. [0016] In another embodiment of the invention, a compound is provided having a formula iv: (R.sub.1)(R.sub.2)(R.sub.3)Si--X-Z-CYNH--Si(NCY).sub.3 iv wherein R.sub.1, R.sub.2 and R.sub.3 independently represents C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; X represents linear or branched C.sub.1-C.sub.20 alkyl or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH, with the proviso that at least one of R.sub.1, R.sub.2, or R.sub.3 represents C.sub.1-C.sub.6 alkoxy. [0017] In another embodiment of the invention, a compound is provided having a formula v: (R.sub.1)(R.sub.2)(R.sub.3)Si--X-Z-CYNH--Si(NHCYL-M).sub.3 v wherein R.sub.1, R.sub.2 and R.sub.3 independently represents C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; X represents linear or branched C.sub.1-C.sub.20 alkyl or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; L represents a linking group; and Z represents oxygen or NH; and M represents a molecule, with the proviso that at least one of R.sub.1, R.sub.2, or R.sub.3 represent C.sub.1-C.sub.6 alkoxy. [0018] In another embodiment of the invention, a compound is provided having a formula vi: ((R.sub.1)(R.sub.2)(R.sub.3)Si--X-Z-CYNH).sub.2--Si(NCY).sub.2 vi wherein R.sub.1, R.sub.2 and R.sub.3 independently represents C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; X represents linear or branched C.sub.1-C.sub.20 alkyl or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH, with the proviso that at least one of R.sub.1, R.sub.2, or R.sub.3 represents C.sub.1-C.sub.6 alkoxy. [0019] In another embodiment of the invention, a compound is provided having a formula vii: ((R.sub.1)(R.sub.2)(R.sub.3)Si--X-Z-CYNH).sub.2--Si(NHCYL-M).sub.2 vii wherein R.sub.1, R.sub.2 and R.sub.3 independently represents C.sub.1-C.sub.6 alkoxy, C.sub.1-C.sub.6 alkyl, phenyl, or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy; L represents a linking group; X represents linear or branched C.sub.1-C.sub.20 alkyl or aryl substituted with one or more groups selected from the group consisting of C.sub.1-C.sub.6 alkyl and C.sub.1-C.sub.6 alkoxy, optionally substituted with one or more heteroatoms comprising oxygen, nitrogen, or sulfur; Y represents oxygen or sulfur; and Z represents oxygen or NH; and M represents a molecule, with the proviso that at least one of R.sub.1, R.sub.2, or R.sub.3 represents C.sub.1-C.sub.6 alkoxy. [0020] In another embodiment of the invention, kits are provided for preparing modified substrates. The kits may include reagents for silylating molecules and optional substrates. [0021] These and other embodiments of the invention will become apparent in light of the detailed description below. Continue reading about Method for attachment of silylated molecules to glass surfaces... Full patent description for Method for attachment of silylated molecules to glass surfaces Brief Patent Description - Full Patent Description - Patent Application Claims Click on the above for other options relating to this Method for attachment of silylated molecules to glass surfaces patent application. ###  How KEYWORD MONITOR works... a FREE service from FreshPatents1. Sign up (takes 30 seconds). 2. Fill in the keywords to be monitored. 3. Each week you receive an email with patent applications related to your keywords. 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