WO1999057323A1 - Chemically modified nucleic acids and methods for coupling nucleic acids to solid support - Google Patents

Chemically modified nucleic acids and methods for coupling nucleic acids to solid support Download PDF

Info

Publication number
WO1999057323A1
WO1999057323A1 PCT/US1999/009810 US9909810W WO9957323A1 WO 1999057323 A1 WO1999057323 A1 WO 1999057323A1 US 9909810 W US9909810 W US 9909810W WO 9957323 A1 WO9957323 A1 WO 9957323A1
Authority
WO
WIPO (PCT)
Prior art keywords
nucleic acid
group
solid support
modified nucleic
modified
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US1999/009810
Other languages
English (en)
French (fr)
Inventor
Allan Bradley
Wei Wen Cai
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Baylor College of Medicine
Original Assignee
Baylor College of Medicine
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Baylor College of Medicine filed Critical Baylor College of Medicine
Priority to CA002326684A priority Critical patent/CA2326684C/en
Priority to EP99920342A priority patent/EP1075544B1/en
Priority to AU37861/99A priority patent/AU770695B2/en
Priority to JP2000547274A priority patent/JP4477774B2/ja
Priority to DE99920342T priority patent/DE99920342T1/de
Priority to AT99920342T priority patent/ATE535615T1/de
Publication of WO1999057323A1 publication Critical patent/WO1999057323A1/en
Anticipated expiration legal-status Critical
Priority to AU2001263094A priority patent/AU2001263094B2/en
Priority to AU2008229872A priority patent/AU2008229872B2/en
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6806—Preparing nucleic acids for analysis, e.g. for polymerase chain reaction [PCR] assay
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H21/00—Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07B—GENERAL METHODS OF ORGANIC CHEMISTRY; APPARATUS THEREFOR
    • C07B2200/00—Indexing scheme relating to specific properties of organic compounds
    • C07B2200/11—Compounds covalently bound to a solid support
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6834—Enzymatic or biochemical coupling of nucleic acids to a solid phase
    • C12Q1/6837—Enzymatic or biochemical coupling of nucleic acids to a solid phase using probe arrays or probe chips
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2525/00—Reactions involving modified oligonucleotides, nucleic acids, or nucleotides
    • C12Q2525/10—Modifications characterised by
    • C12Q2525/197—Modifications characterised by incorporating a spacer/coupling moiety
    • C—CHEMISTRY; METALLURGY
    • C40—COMBINATORIAL TECHNOLOGY
    • C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
    • C40B40/00—Libraries per se, e.g. arrays, mixtures

Definitions

  • the present Invention claims a closely related family of compounds, devices, and methods relating to techniques for immobilizing nucleic acids to a solid support for the purpose of conducting scientific investigation or routine testing upon the bound nucleic acid samples in areas such as genome-wide genetic mapping and gene expression studies.
  • 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.
  • immobilizing DNA means fixing one end of the strand to the solid support so that the remainder of the strand is unmodified and free to undergo further reaction depending upon the particular study. Indeed, this is a widely used method to conduct laboratory studies involving DNA.
  • the particular immobilized strand to which the probe reacts reveals the nucleotide sequence of the previously unknown immobilized strand.
  • probe studies are severely confounded by electrostatic sticking of the probe to the derivatized (hence electrostatically charged) glass surface.
  • the probe is often radiolabeled so that its presence can be detected by an ordinary radiation detector.
  • the location of the probe on the glass surface as evidenced by the detector, reveals the chemical identity or sequence of the immobilized DNA strand at that particular location on the glass surface (which is known and designated in advance) .
  • 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) .
  • One very common substance used to prepare a glass surface to receive a nucleic acid sample is poly-L- lysine. See, e.g., DeRisi, et al . , Use of a cDNA Microarray to Analyze Gene Expression Pattern in Human Cancer, 14 Nature Genetics 457 (1996); Shalon et al . in A DNA Microarray System for Analyzing Complex DNA Samples Using Two -Color Fluorescent Probe Hybridization, 6 Genome Res. 639 (1996); and Schena, et al . , Quanti tative Moni toring of Gene Expression Patterns Wi th a Complementary DNA Microarray, 270 Science 467 (1995) .
  • pre-derivatized glass supports are commercially available (e.g., silylated microscope slides). See, e.g., Schena, et al . , Parallel Human Genome Analysis : Microarray-Based Expression Moni toring of 1000 Genes, 93 P.N.A.S. 10614 (1996) .
  • U.S. Pat. No. 5,630,932 assigned to Molecular Imaging Corp. discloses a coating for a probe (platinum) tip for use in scanning tunneling microscopy; numerous means are disclosed for coating the surface, notably, Si(OCH 3 )CH 2 I.
  • U.S. Pat. No. 5,610,287, assigned to Molecular Tool discloses coating a solid support with a salt or cationic detergent to non-covalently bond nucleic acids to the support.
  • U.S. Pat. No. 5,024,933, assigned to Enzo Biochem discloses coating a solid support with an isolate of naturally occurring mussel adhesive protein.
  • Another approach to this problem involves derivatizing both the solid support and the nucleic acid sought to be immobilized. See, e.g., U.S. Pat. No. 5,641,630, assigned to Amgen and Abbott, discloses coating a solid support with a complexing agent that binds to an other complexing agent to which the nucleic acid sought to be bound is likewise bound.
  • U.S. Pat No. 5,554,744, assigned to Hybridon discloses contacting a solid support with diisopropylcarbodiimide and an acid catalyst and a succinylated nucleoside to immobilize the nucleoside.
  • 5,514,785 assigned to Becton Dickinson, discloses coating a solid support with, preferably, primary and secondary amines, followed by activation of the nucleic acid using cyanuric chloride.
  • U.S. Pat. No. 5,215,882, assigned to Ortho Diagnostic Systems discloses modifying the nucleic acid sought to be immobilized with a primary amine or equivalent, followed by reaction of the modified nucleic acid with the solid support (the support must have free aldehyde groups) in the presence of a reducing agent.
  • the prevailing view in the biochemical arts is that, in order to effectively immobilize nucleic acids onto solid surfaces, the solid support must first be derivatized, or made chemically labile, so that the nucleic acid can then be reacted with solid support.
  • epoxides are known mutagens; that is, they are known to damage nucleic acids, particularly DNA. Therefore, quite contrary to the current state of knowledge in the biochemical arts, the Invention presented here discloses and claims DNA (and nucleic acid more generally) that is modified such that they readily adhere to an unmodified or underivatized glass surface.
  • the present Invention discloses and claims epoxide -modified nucleic acid (particularly DNA) which is readily affixed to an unmodified solid support.
  • One object of the present Invention is modified nucleic acid that will adhere to a solid surface to allow subsequent biochemical investigation.
  • a modified nucleic acid which comprises a nucleic acid covalently bound to moiety containing two crucial functional groups: a cyclic ether group and an alkoxysilane group.
  • methods for preparing the aforementioned modified nucleic acids are claimed.
  • a high-density microarray which comprises a glass or other inert surface, made by printing numerous highly discrete modified DNA sample spots upon the surface.
  • another modified nucleic acid is claimed which is prepared from a nucleic acid and a halogenated silane .
  • nucleic acid is claimed which is prepared by reaction of the nucleic acid with a brominated moiety, followed by reaction with an a inated silane .
  • a device which allows printing of the aforementioned high-density microarrays.
  • modified silanes are claimed which allow the skilled artisan to modulate the electrostatic properties of the solid surface to optimize sample density and detection sensitivity.
  • 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
  • the nucleic acid to be immobilized upon the solid support is readily derivatized.
  • the reaction of the epoxide derivatives of the present Invention is simply to execute—it occurs under mild conditions, reaction rates are quick, and equilibrium is highly favorable.
  • the epoxide-modified nucleic acid 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.
  • Figure 1 depicts a coupling reaction of nucleic acid (in this instance DNA) with 3- glycidoxypropyltrimethoxysilane, followed by the reaction of the newly modified DNA and the solid support (in this instance a glass surface) .
  • the final reaction product—the immobilized DNA is shown at bottom.
  • Figure 2 depicts a coupling reaction of nucleic acid
  • Figure 3 depicts a device for making a high-density microarray; both a top (Fig. 3A) and a side view (Fig. 3B) are shown.
  • Figure 4 depicts the silanization of nucleic acid through alkylation of halogen-containing silane compounds .
  • Figure 5a depicts the first step in the silanization of nucleic acid using amine-containing silane compounds. In this case, the reaction occurs preferentially at the guanine base at neutral and slightly basic pH.
  • Figure 5b depicts the first step in the silanization of nucleic acid using amine-containing silane compounds. In this case, the reaction occurs preferentially at the cytosine base at more basic pH.
  • Figure 5c depicts the second and final step in the silanization of nucleic acid using amine-containing silane compounds.
  • Figure 6 is a schematic representation of one embodiment of the present Invention showing silane linkers by hydrophobic linkers.
  • the gist of this invention is chemical modification of the nucleic acid sought to be immobilized. This chemically modified nucleic acid is then readily reacted to a solid support such as a glass surface, rendering the nucleic acid immobilized. Again, this is in direct contradiction to the prior art, which teaches modification of the solid support, rather than the nucleic acid itself.
  • the modified nucleic acids of the present Invention readily adhere to a variety of solid surfaces having hydroxyl groups. These include, though are not limited to: quartz glass, mica, alumina (A1 2 0 3 ) , titania (Ti0 2 ) , Sn0 2 , Ru0 2 , Pt0 2 , as well as numerous other metal oxide surfaces .
  • the chemically modified nucleic acids of the present Invention are so modified with compounds having two crucial functionalities: a ring ether and an alkoxysilane group. The nucleic acid reacts with the ring ether, then the newly modified nucleic acid is contacted with the otherwise inert glass surface, where the alkoxysilane group reacts with the Si-OH groups on the glass surface.
  • the chemically modified nucleic acids of the present Invention are so modified with compounds having two crucial functionalities: an amino group and an alkoxysilane group.
  • the nucleic acid reacts with the amino group, then the newly modified nucleic acid is contacted with the otherwise inert glass surface, where the alkoxysilane group reacts with the Si-OH groups on the glass surface.
  • nucleic acids are modified by reaction with halogenated
  • the nucleic acids are derivatized by a two-step process involving a final reaction with amine-containing silanes and brominated nucleic acids.
  • Other embodiments are directed to preparing and optimizing high-density microarrays utilizing the modified nucleic acids of the prior embodiments of the present Invention.
  • nucleic acid of the present Invention describes one form of modified nucleic acid of the present Invention.
  • the purpose of the chemical modification is to enable the nucleic acid to be readily affixed to an underivatized solid surface.
  • the nucleic acid preferably DNA
  • the nucleic acid is modified by reaction with 3 - glycidoxypropyltrimethoxysilane (GPTS) , according to Fig. 1.
  • GPTS 3 - glycidoxypropyltrimethoxysilane
  • GPTS 3 glycidoxypropyltrimethoxysilane
  • GPTS 3 glycidoxypropyltrimethoxysilane
  • affixing the nucleic acid to the solid support consists essentially of two steps.
  • the nucleic acid reacts with the epoxide end of the GPTS molecule; in the second step, the glass surface
  • the compound shown is 3-glycidoxypropyltrimethoxysilane or GPTS.
  • DNA is reacted with GPTS at basic pH, preferably above 9.5, to form the modified DNA.
  • the modified DNA is then reacted with an underivatized glass (or other silanol-containing) surface at neutral pH, thus immobilizing the DNA onto the glass surface.
  • the ring ether functionality reacts with the DNA.
  • the ring ether need not be ethylene oxide, as it is in GPTS, although the small ring is preferred to increase reactivity of the ether functionality which is relatively unreactive.
  • the first reaction leading to the derivatized DNA, is a ring-opening reaction likely involving carbon 5 of the ribose ring of the DNA.
  • This derivatized DNA is unusually stable and can be stored for long periods of time prior to actual use.
  • the second reaction, immobilizing the derivatized DNA onto the glass surface, is a simple substitution reaction creating an Si-O-Si
  • nucleic acid of the present Invention describes another preferred form of modified nucleic acid of the present Invention.
  • the purpose of the chemical modification is to enable the nucleic acid to be readily affixed to an underivatized solid surface.
  • the nucleic acid preferably DNA
  • the nucleic acid is modified by reaction with 3- aminopropyltrimethoxysilane, according to Fig. 2.
  • affixing the nucleic acid to the solid support consists essentially of two steps.
  • the nucleic acid reacts with the epoxide end of the 3- aminopropyltrimethoxysilane molecule; in the second step, the glass surface reacts with the other end, or the silane end of the 3-aminopropyltrimethoxysilane-modified nucleic acid, thereby affixing the nucleic acid onto an underivatized glass surface.
  • the entire reaction is rapid, is characterized by a favorable equilibrium, and occurs under very mild conditions using a minimum of inexpensive reagents. Though there quite obviously are numerous ways to carry out either step of the reaction, the preferred method is shown in this and the following example.
  • the compound shown is 3- aminopropyltriethoxysilane .
  • the first reaction, leading to the derivatized DNA, is transamination reaction of the cytosine residues on nucleic acids.
  • the second reaction as in Example 1, immobilizing the derivatized DNA onto the glass surface is a simple substitution reaction creating an Si-O-Si linkage in the glass surface, and removing one of the alkoxy groups from the GPTS molecule.
  • modified nucleic acids of the present Invention such as those described in Examples 1 and 2
  • these modified nucleic acids can be immobilized onto a glass surface simply by contacting the modified DNA onto the underivatized surface.
  • the significance of this is, among other things, that spreading (migration of the DNA sought to be immobilized from the desired location) and non-specific probe sticking (caused by derivatization of the glass surface which creates a net positive electrostatic charge upon the surface which attracts the net negatively charged DNA) are essentially eliminated.
  • a high-density microarray consisting of multiple DNA samples of this type is also easily constructed in accordance with the present Invention.
  • the modified DNA can be prepared (for instance, in accordance with Examples 1 and 2) well in advance of actual use.
  • These chemically modified DNA samples are analogous to "DNA chips" that can then be readily “imprinted” upon an unaltered glass sheet in, for instance, grid fashion.
  • Fig. 3 illustrates one embodiment of a device for preparing such a high-density microarray using the DNA chips of the present Invention.
  • the device is made from a plurality of inexpensive commercially available capillary micropipets, preferably 10 cm micropipets, although other sizes will, of course, work. As depicted in Fig.
  • each 10 cm micropipet is pulled to make a taper at one end. They are arranged in a hexagonal close-packed array, bounded by a square frame. The micropipets can be glued to one another to form a stable unit within the frame . The tapered ends (Fig. 3B) are cut off and polished to optical flatness.
  • the tips of the device are dipped into a multi-well container which contains the (chemically modified in accordance with the present Invention) DNA samples to be tested, and whose wells are aligned with the micropipets of the device.
  • a small portion of each DNA sample is deposited into the micropipet corresponding to the particular well by simple capillary action.
  • the size of the spot can be carefully controlled by the size of the tapered end.
  • the present Invention thousands of samples can be arrayed in a narrow area, simultaneously and without the need for expensive robotics.
  • the method (comprising the DNA chips and pipet device) of the present Invention has been shown to be even more efficient than methods using high-speed spotting robots.
  • the compounds, methods and devices of the present Invention are readily incorporated into a prepackaged kit for commercial sale.
  • the high-density microarray of the present Invention can also be readily incorporated into the microarray systems of the prior art, such as those disclosed in the prior art section above. These methods are hereby incorporated by reference into the present Application, for instance, fluorescent in si tu hybridization (FISH) and the method described in Shalon, et al .
  • FISH fluorescent in si tu hybridization
  • a modified nucleic acid in accordance with the present Invention is prepared by reacting unmodified nucleic acid under near neutral pH with suitable silane compounds.
  • the "X" in Fig. 4 can refer to any halide, preferably Cl , Br, or I; R lf R 2 , and R 3 , can be the same or different, including, --OCH 3 , and --OC 2 H 5 .
  • the halogenated silane depicted to the left of the arrow in Fig. 4 is 8- bromocytltrichlorosilane, 8-bromocytltrimethoxysilane, 4-chlorobutylmethyldichlorosilane, and 3- iodopropyltrimethoxysilane .
  • the conversion depicted in Fig. 4 was performed as follows.
  • the halogenated silane was dissolved in dimethylformamide (DMF) at a concentration of about 30 ⁇ iM.
  • 3 to 10 ug of nucleic acid was dissolved in 100 ul of 0.01 M phosphate buffer (pH 7.0) .
  • 1 to 3 ug of 30 mM halogenated silane was added, the solution is then mixed well, and allowed to react at about 37 C for about 3 hours (alternatively, it can be reacted at ambient temperature overnight) .
  • the desired product the modified nucleic acid—is purified by ethanol precipitation; then the modified nucleic acid is dissolved in water.
  • one particular advantage of the present Invention is that it allows the investigator to prepare unusually high-density microarrays to conduct nucleic acid studies.
  • This example is best understood in relation to example 3 which disclosed the preparation of a high-density microarray in accordance with the present Invention.
  • This example discloses enhanced methods for controlling the size of the individual nucleic acid "spots" on the solid supports, in accordance with the present Invention.
  • silanes of the present Invention are quite general embodiments of these silanes after hydrolysis contains an Si (OH) 3 at each end, linked by a hydrophobic group. See Figure 6. Any of a variety of hydrophobic linkers can be used. Particularly preferred embodiments include: 1,6-Bis- trichlorosilyhexane, 1 , 8-Bis-trichlorosilyloctane, 1,6- Bis-trimethoxysilyhexane, and 1,4 Bis- trimethoxysilylethylbenzene .
  • one end of the silane attaches to the surface, and the other end remains reactive to the modified nucleic acids.
  • the hydrophobic linker confers hydrophobicity to the surface.
  • the skilled artisan can readily see how the electrostatic properties of the surface (hydrophobic versus hydrophilic) can be readily modulated—e .g. , the chain length of the linker can be adjusted to control hydrophobicity, and the surface reactivity can be controlled by adjusting the amount of silane contacted with the surface.
  • the glass surface was cleaned by slowly boiling in 3 M HCl for about 2 hrs in a fume hood. Next, the surfaces were rinsed with deionized water then kept in 0.1 M HCl until ready for use. When ready for use, the surfaces were rinsed with doubly distilled deionized water to remove any extant acid, then rinsed in absolute ethanol . Next, the surfaces were immediately transferred to an ethanol solution containing 0.0005 % to 0.002 % of the bi- functional silanes of this aspect of the Invention. The surfaces were then treated at room temperature for
  • the modified nucleic acid is prepared by reacting pristine nucleic acids with an amine-containing silane.
  • the derivatization of nucleic acid with amine-containing silanes is comprised of two steps: (1) the halogenation (or bromination, as shown) of the nucleic acid (Fig. 5a, 5b) ; and (2) the derivatization of the halogenated nucleic acid (Fig. 5c) .
  • the reaction can occur in the presence of N-bromosuccinimide under mild pH conditions; varying either of these reaction variables allows the skilled biochemist to control the reaction rate.
  • the reaction normally occurs at the guanine or cytosine base depending upon the pH—i . e . , neutral to slightly basic pH favors reaction at the guanine residue, more basic pH favors reaction at the cytosine residue.
  • nucleic acid is DNA or RNA.
  • DNA 5 ug of DNA was dissolved in lOO ul of O.l M NaHC0 3 , to reach a pH of about 9.5. This solution is kept on ice for about 5 minutes. Contemporaneously, a fresh N-bromosuccinimide solution
  • -20- at concentration of about 10 mM was prepared and also chilled on ice.
  • 1 ul of the N-bromosuccinimide solution is added to the DNA solution; the solution was then stirred vigorously (to vortex) . The reaction was then allowed to proceed on ice for about 15 minutes.
  • 10 ul of 0.5 M aminosilane solution at pH about 9.5 - 12 was added to the bromine-activated DNA solution; this new mixture was allowed to react at 65 C for about 2 hours.
  • the silane-modified DNA was purified by methods well known in the art; preferably, it is purified by ethanol precipitation.
  • RNA was dissolved in 100 ul of 0.1 M phosphate buffer, to reach a pH of about 7.5. This solution is kept on ice for about 5 minutes.
  • N-bromosuccinimide solution at concentration of about 10 mM was prepared and also chilled on ice.
  • 1 ul of the N-bromosuccinimide solution is added to the DNA solution; the solution was then stirred vigorously (to vortex) . The reaction was then allowed to proceed on ice for about 15 minutes.
  • 10 ul of 1 M aminosilane solution at pH about 8.0 was added to the bromine-activated DNA solution; this new mixture was allowed to react at 45 C for about 2 hours.
  • the silane-modified DNA was purified by methods well known in the art; preferably, it is purified by ethanol precipitation.
  • H 2 N-X- Si -OR R2 H , -CH 3 , -C 2 H 5 , -OCH 3 , -OC 2 H 5

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Biophysics (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Saccharide Compounds (AREA)
PCT/US1999/009810 1998-05-04 1999-05-04 Chemically modified nucleic acids and methods for coupling nucleic acids to solid support Ceased WO1999057323A1 (en)

Priority Applications (8)

Application Number Priority Date Filing Date Title
CA002326684A CA2326684C (en) 1998-05-04 1999-05-04 Chemically modified nucleic acids and methods for coupling nucleic acids to solid support
EP99920342A EP1075544B1 (en) 1998-05-04 1999-05-04 Chemically modified nucleic acids and methods for coupling nucleic acids to solid support
AU37861/99A AU770695B2 (en) 1998-05-04 1999-05-04 Chemically modified nucleic acids and methods for coupling nucleic acids to solid support
JP2000547274A JP4477774B2 (ja) 1998-05-04 1999-05-04 化学修飾された核酸および核酸の固体支持体への結合方法
DE99920342T DE99920342T1 (de) 1998-05-04 1999-05-04 Chemisch modifizierte Nukleinsäuren und Methode zur Kopplung von Nukleinsäuren an einen festen Träger
AT99920342T ATE535615T1 (de) 1998-05-04 1999-05-04 Chemisch modifizierte nukleinsäuren und methode zur kopplung von nukleinsäuren an einen festen träger
AU2001263094A AU2001263094B2 (en) 1998-05-04 2001-05-10 Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
AU2008229872A AU2008229872B2 (en) 1998-05-04 2008-10-10 Compositions and methods for array-based genomic nucleic acid analysis of biological molecules

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US09/071,876 US6048695A (en) 1998-05-04 1998-05-04 Chemically modified nucleic acids and methods for coupling nucleic acids to solid support
US09/071,876 1998-05-04

Publications (1)

Publication Number Publication Date
WO1999057323A1 true WO1999057323A1 (en) 1999-11-11

Family

ID=22104163

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US1999/009810 Ceased WO1999057323A1 (en) 1998-05-04 1999-05-04 Chemically modified nucleic acids and methods for coupling nucleic acids to solid support

Country Status (8)

Country Link
US (3) US6048695A (https=)
EP (1) EP1075544B1 (https=)
JP (1) JP4477774B2 (https=)
AT (1) ATE535615T1 (https=)
AU (1) AU770695B2 (https=)
CA (1) CA2326684C (https=)
DE (1) DE99920342T1 (https=)
WO (1) WO1999057323A1 (https=)

Cited By (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1006363A3 (en) * 1998-12-01 2000-08-16 Hitachi Software Engineering Co., Ltd. Biochip and method for producing the same
WO2001042501A1 (en) * 1999-12-09 2001-06-14 Karolinska Innovations Ab Method for immobilisation
DE19957827A1 (de) * 1999-11-25 2001-06-21 Epigenomics Ag Oligomer-Array mit PNA- und/oder DNA-Oligomeren auf einer Oberfläche
EP1110967A1 (en) * 1999-12-21 2001-06-27 LION Bioscience AG Compound comprising a biomolecule moiety and an organo-silane moiety
WO2001046214A3 (en) * 1999-12-21 2001-12-27 Lion Bioscience Ag Compound comprising a nucleic acid moiety and an organo-silane moiety
WO2001046461A3 (en) * 1999-12-22 2002-05-23 Biochip Technologies Gmbh Modified nucleic acids and their use
WO2002092615A3 (en) * 2001-05-10 2002-12-19 Baylor College Medicine Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
DE10139283A1 (de) * 2001-08-09 2003-03-13 Epigenomics Ag Verfahren und Nukleinsäuren zur Analyse von Colon-Krebs
JP2003520350A (ja) * 2000-01-20 2003-07-02 サントル・ナショナル・ドゥ・ラ・ルシェルシュ・シアンティフィク−シーエヌアールエス シラン化固体支持体上で核酸を合成および固定化するための方法
JP2003521680A (ja) * 2000-01-11 2003-07-15 ナノゲン・レコグノミクス・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツング 基体表面に結合させるための複数の結合部分を有する生体分子
JP2004534959A (ja) * 2001-07-09 2004-11-18 サントル　ナショナル　ドゥ　ラ　ルシェルシュ　シアンティフィク 固形支持体の機能化方法、機能化された固形支持体、およびその使用
US6858713B1 (en) 1998-05-04 2005-02-22 Baylor College Of Medicine Chemically modified biological molecules and methods for coupling biological molecules to solid support
US6979728B2 (en) 1998-05-04 2005-12-27 Baylor College Of Medicine Articles of manufacture and methods for array based analysis of biological molecules
WO2006072045A3 (en) * 2004-12-30 2006-08-24 Corning Inc Substrates having pendant epoxide groups for binding biomolecules and methods of making and using thereof
US7195908B2 (en) 2002-10-31 2007-03-27 Corning Incorporated Supports treated with triamine for immobilizing biomolecules
US7217512B2 (en) 2002-05-09 2007-05-15 Corning Incorporated Reagent and method for attaching target molecules to a surface
EP1451318A4 (en) * 2001-10-12 2007-06-27 Perkinelmer Las Inc COMPOSITIONS OF NUCLEIC ACIDS AND ARRAYS AND METHOD FOR THEIR USE
AU2001263094B2 (en) * 1998-05-04 2008-07-10 Baylor College Of Medicine Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
DE102008053270A1 (de) * 2008-10-27 2010-05-12 Medizinische Hochschule Hannover Vorrichtung und Verfahren zur Analyse von Zellen
FR3038734A1 (fr) * 2015-07-10 2017-01-13 Centre Nat De La Rech Scient (Cnrs) Nouveaux materiaux optiques fonctionnalises
US9671396B2 (en) * 2001-09-05 2017-06-06 Joon Won Park Solid substrate comprising array of dendrons and methods for using the same

Families Citing this family (160)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6426183B1 (en) * 1995-12-21 2002-07-30 Kenneth L. Beattie Oligonucleotide microarrays: direct covalent attachment to glass
US7078224B1 (en) 1999-05-14 2006-07-18 Promega Corporation Cell concentration and lysate clearance using paramagnetic particles
US6248127B1 (en) * 1998-08-21 2001-06-19 Medtronic Ave, Inc. Thromboresistant coated medical device
JP2001149060A (ja) * 1999-11-29 2001-06-05 Nisshinbo Ind Inc 核酸固定化基板
AU6291301A (en) * 2000-02-22 2001-09-03 Genospectra, Inc. Microarray fabrication techniques and apparatus
CN1404415A (zh) 2000-02-22 2003-03-19 基因谱公司 微阵列制造技术及设备
US20040014102A1 (en) * 2000-02-22 2004-01-22 Shiping Chen High density parallel printing of microarrays
JP3502803B2 (ja) * 2000-03-06 2004-03-02 日立ソフトウエアエンジニアリング株式会社 マイクロアレイ、マイクロアレイ作製方法及びマイクロアレイにおけるピン間スポット量誤差補正方法
WO2001075166A2 (en) * 2000-03-31 2001-10-11 Genentech, Inc. Compositions and methods for detecting and quantifying gene expression
KR100352171B1 (ko) * 2000-04-14 2002-09-12 (주) 제노텍 올리고뉴클레오타이드를 지지체에 고정시키는 방법 및 그방법에 의하여 제조되는 올리고뉴클레오타이드 어레이
BR0106969A (pt) * 2000-07-05 2004-07-13 Cuno Inc Método de fabricação de um substrato multicelular e substrato multicelular
US6890483B2 (en) 2000-07-05 2005-05-10 Cuno Incorporated Non-luminescent substrate
US20030219816A1 (en) * 2001-07-02 2003-11-27 Keith Solomon Composite microarray slides
JP2004506201A (ja) * 2000-08-03 2004-02-26 マサチューセッツ・インスティチュート・オブ・テクノロジー 機能性生体分子のマイクロアレイおよびその使用
US20020102617A1 (en) * 2000-08-03 2002-08-01 Macbeath Gavin Protein microarrays
KR100379720B1 (ko) * 2000-10-14 2003-04-11 주식회사 마크로젠 덴드리머 단일층 지지체 및 그의 제조방법
EP1334113A4 (en) * 2000-10-20 2007-08-08 Expression Diagnostics Inc EVALUATION OF LEUCOCYTAIRE EXPRESSION LEVEL
US6861214B1 (en) * 2000-10-23 2005-03-01 Beckman Coulter, Inc. Immobilization of biopolymers to aminated substrates by direct adsorption
US20020150887A1 (en) * 2000-11-09 2002-10-17 National Institute Of Advanced Industrial Science And Technology Methods and nucleic acid probes for molecular genetic analysis of polluted environments and environmental samples
US20020146684A1 (en) * 2001-04-09 2002-10-10 Meldal Morten Peter One dimensional unichemo protection (UCP) in organic synthesis
JP2004530879A (ja) * 2001-05-03 2004-10-07 シグマ−ジェノシス リミテッド タンパク質マイクロアレイを構築する方法
US7235358B2 (en) 2001-06-08 2007-06-26 Expression Diagnostics, Inc. Methods and compositions for diagnosing and monitoring transplant rejection
US6905827B2 (en) * 2001-06-08 2005-06-14 Expression Diagnostics, Inc. Methods and compositions for diagnosing or monitoring auto immune and chronic inflammatory diseases
US7026121B1 (en) 2001-06-08 2006-04-11 Expression Diagnostics, Inc. Methods and compositions for diagnosing and monitoring transplant rejection
US10272409B2 (en) 2001-07-11 2019-04-30 Michael E. Hogan Methods and devices based upon a novel form of nucleic acid duplex on a surface
CA2656209C (en) * 2001-07-11 2012-11-27 Baylor College Of Medicine Methods and devices based upon a novel form of nucleic acid duplex on a surface
US20030087309A1 (en) * 2001-08-27 2003-05-08 Shiping Chen Desktop drug screening system
US20050032060A1 (en) * 2001-08-31 2005-02-10 Shishir Shah Arrays comprising pre-labeled biological molecules and methods for making and using these arrays
WO2003027638A2 (en) * 2001-09-27 2003-04-03 Spectral Genomics, Inc. Methods for detecting genetic mosaicisms using arrays
US7439346B2 (en) * 2001-10-12 2008-10-21 Perkinelmer Las Inc. Nucleic acids arrays and methods of use therefor
US20030124599A1 (en) * 2001-11-14 2003-07-03 Shiping Chen Biochemical analysis system with combinatorial chemistry applications
US20030157527A1 (en) * 2001-12-03 2003-08-21 Invitrogen Corporation Identification of rearrangements in nucleic acid molecules
WO2003056007A1 (fr) * 2001-12-26 2003-07-10 Canon Kabushiki Kaisha Materiau de sonde
KR100450191B1 (ko) * 2001-12-28 2004-10-02 삼성에스디아이 주식회사 생체물질 고정용 기판 및 이의 제조방법
WO2003076903A2 (en) * 2002-03-08 2003-09-18 Spectral Genomics, Inc. Articles of manufacture and methods for making hydrophobic derivatized arrays
US6916621B2 (en) * 2002-03-27 2005-07-12 Spectral Genomics, Inc. Methods for array-based comparitive binding assays
US7226771B2 (en) 2002-04-19 2007-06-05 Diversa Corporation Phospholipases, nucleic acids encoding them and methods for making and using them
PT1497418E (pt) 2002-04-19 2013-01-25 Dsm Ip Assets Bv Fosfolípases, ácidos nucleicos que as codificam e métodos para as preparar e utilizar
AU2003272548A1 (en) * 2002-09-16 2004-04-30 Plexxikon, Inc. Crystal structure of pim-1 kinase
US7129046B2 (en) * 2002-10-21 2006-10-31 Agilent Technologies, Inc. Linking to chemical array assemblies with metal layers
US20050048573A1 (en) * 2003-02-03 2005-03-03 Plexxikon, Inc. PDE5A crystal structure and uses
JP2007524374A (ja) * 2003-02-28 2007-08-30 プレキシコン，インコーポレーテッド Ｐｙｋ２結晶構造および使用
CN102618564B (zh) 2003-03-06 2014-10-29 维莱尼姆公司 淀粉酶、编码它们的核酸及其制备和应用方法
CA3007908A1 (en) 2003-03-07 2005-04-14 Dsm Ip Assets B.V. Hydrolases, nucleic acids encoding them and methods for making and using them
HUE030493T2 (en) 2003-04-04 2017-05-29 Basf Enzymes Llc Pectate lyases, nucleic acids and processes encoding them for their production and use
US7892745B2 (en) 2003-04-24 2011-02-22 Xdx, Inc. Methods and compositions for diagnosing and monitoring transplant rejection
US20070248978A1 (en) * 2006-04-07 2007-10-25 Expression Diagnostics, Inc. Steroid responsive nucleic acid expression and prediction of disease activity
FR2854696A1 (fr) * 2003-05-06 2004-11-12 Commissariat Energie Atomique Support de biopuce utilisant des couches minces de materiau sol gel et procede de realisation
US7960148B2 (en) 2003-07-02 2011-06-14 Verenium Corporation Glucanases, nucleic acids encoding them and methods for making and using them
US20050079548A1 (en) * 2003-07-07 2005-04-14 Plexxikon, Inc. Ligand development using PDE4B crystal structures
CA2535526C (en) 2003-08-11 2015-09-29 Diversa Corporation Laccases, nucleic acids encoding them and methods for making and using them
WO2005028624A2 (en) * 2003-09-15 2005-03-31 Plexxikon, Inc. Molecular scaffolds for kinase ligand development
US7279280B2 (en) * 2003-09-25 2007-10-09 Mgp Biotech, Inc. Apparatus and method for detecting genetic mutations and single nucleotide polymorphisms
DK2322278T3 (en) 2003-10-24 2017-04-10 Aushon Biosystems Inc Apparatus and method for dispensing liquid, semi-solid and solid samples
EP1678329A4 (en) * 2003-10-30 2008-07-02 Tufts New England Medical Ct PRENATAL DIAGNOSIS USING CELL-FREE FEDERAL DNA IN FRUIT WATER
US20070066641A1 (en) * 2003-12-19 2007-03-22 Prabha Ibrahim Compounds and methods for development of RET modulators
CN1925855B (zh) * 2003-12-19 2010-06-16 普莱希科公司 开发Ret调节剂的化合物和方法
JP4202391B2 (ja) * 2004-03-17 2008-12-24 パナソニック株式会社 バイオチップの製造方法、プローブ溶液、および、バイオチップ
EP1742627A4 (en) 2004-05-06 2009-08-26 Plexxikon Inc PDE4B HEMMER AND ITS USE
US8911942B2 (en) * 2004-05-20 2014-12-16 Quest Diagnostics Investments Incorporated Single label comparative hybridization
AU2005264938B2 (en) 2004-06-16 2011-11-24 Verenium Corporation Compositions and methods for enzymatic decolorization of chlorophyll
JP2008503473A (ja) * 2004-06-17 2008-02-07 プレキシコン，インコーポレーテッド Ｃ−ｋｉｔ活性を調節する化合物
US7498342B2 (en) 2004-06-17 2009-03-03 Plexxikon, Inc. Compounds modulating c-kit activity
WO2006026754A2 (en) * 2004-09-03 2006-03-09 Plexxikon, Inc. Bicyclic heteroaryl pde4b inhibitors
WO2006029184A2 (en) * 2004-09-08 2006-03-16 Expression Diagnostics, Inc. Genes useful for diagnosing and monitoring inflammation related disorders
US20090142854A1 (en) * 2004-11-16 2009-06-04 Robert Veronique Silanizing agents comprising a saccharide end group and uses thereof, in particular for the functionalization of solid supports
WO2006060742A2 (en) * 2004-12-02 2006-06-08 Oncotech, Inc. Reagents and methods for predicting drug resistance
US8021888B2 (en) * 2005-01-27 2011-09-20 Quest Diagnostics Investments Incorporated Rapid comparative genomic hybridization using acoustic surface waves
EP2886658A1 (en) 2005-03-10 2015-06-24 BASF Enzymes LLC Lyase enzymes, nucleic acids encoding them and methods for making and using them
CA2611859C (en) 2005-03-15 2015-03-31 Verenium Corporation Cellulases, nucleic acids encoding them and methods for making and using them
WO2006102497A2 (en) * 2005-03-22 2006-09-28 The Board Of Trustees Of The Leland Stanford Junior University Methods and compositions for diagnosis, monitoring and development of therapeutics for treatment of atherosclerotic disease
EP1885889A4 (en) * 2005-05-11 2010-01-20 Expression Diagnostics Inc METHOD FOR MONITORING THE OPERATING STATUS OF TRANSPLANTS OVER GENLISTS
US7846941B2 (en) * 2005-05-17 2010-12-07 Plexxikon, Inc. Compounds modulating c-kit and c-fms activity and uses therefor
UA95244C2 (ru) * 2005-06-22 2011-07-25 Плексикон, Инк. Соединения и способ модулирования активности киназ, и показания для их применения
US20070015191A1 (en) * 2005-07-01 2007-01-18 Promega Corporation Network of buoyant particles for biomolecule purification and use of buoyant particles or network of buoyant particles for biomolecule purification
CN1908189A (zh) * 2005-08-02 2007-02-07 博奥生物有限公司 体外辅助鉴定肠型胃癌及其分化程度的方法与专用试剂盒
US8076074B2 (en) 2005-11-29 2011-12-13 Quest Diagnostics Investments Incorporated Balanced translocation in comparative hybridization
EP1963526A4 (en) 2005-12-09 2009-11-18 Promega Corp PURIFICATION OF NUCLEIC ACID WITH A BINDING MATRIX
US20100009373A1 (en) * 2005-12-23 2010-01-14 Perkinelmer Health Sciences, Inc. Methods and compositions relating to multiplex genomic gain and loss assays
US20090104613A1 (en) * 2005-12-23 2009-04-23 Perkinelmer Las, Inc. Methods and compositions relating to multiplexed genomic gain and loss assays
US20070166739A1 (en) 2005-12-23 2007-07-19 Perkinelmer Las, Inc. Comparative genomic hybridization on encoded multiplex particles
US7932037B2 (en) 2007-12-05 2011-04-26 Perkinelmer Health Sciences, Inc. DNA assays using amplicon probes on encoded particles
EP1987142A4 (en) 2006-02-02 2009-07-15 Verenium Corp ESTERASES AND ASSOCIATED NUCLEIC ACIDS AND METHODS
NZ595497A (en) 2006-02-10 2013-09-27 Verenium Corp Cellulolytic enzymes, nucleic acids encoding them and methods for making and using them
TR201809912T4 (tr) 2006-02-14 2018-07-23 Bp Corp North America Inc Ksilanazlar, bunları kodlayan nükleik asitler ve bunları yapmak ve kullanmak için yöntemler.
MX2008011477A (es) 2006-03-07 2008-09-23 Cargill Inc Aldolasas, acidos nucleicos que las codifican y metodos para hacerlas y usarlas.
AU2007223086B2 (en) 2006-03-07 2013-09-26 Basf Enzymes Llc Aldolases, nucleic acids encoding them and methods for making and using them
US20070232556A1 (en) * 2006-03-31 2007-10-04 Montine Thomas J Methods and compositions for the treatment of neurological diseases and disorders
DK2069389T3 (en) 2006-08-04 2015-01-12 Bp Corp North America Inc Glucanases, nucleic acids encoding them, and processes for their preparation and use
WO2008021431A2 (en) * 2006-08-14 2008-02-21 Xdx, Inc. Methods and compositions for diagnosing and monitoring the status of transplant rejection and immune disorders
ES2531135T3 (es) 2006-09-21 2015-03-11 Basf Enzymes Llc Fitasas, ácidos nucleicos que las codifican y métodos para su producción y uso
EP2057274B1 (en) 2006-09-21 2013-12-11 DSM IP Assets B.V. Phospholipases, nucleic acids encoding them and methods for making and using them
US8618248B2 (en) 2006-10-31 2013-12-31 President And Fellows Of Harvard College Phosphopeptide compositions and anti-phosphopeptide antibody compositions and methods of detecting phosphorylated peptides
EP2102367A2 (en) * 2006-11-09 2009-09-23 XDX, Inc. Methods for diagnosing and monitoring the status of systemic lupus erythematosus
WO2008063888A2 (en) 2006-11-22 2008-05-29 Plexxikon, Inc. Compounds modulating c-fms and/or c-kit activity and uses therefor
WO2008079909A1 (en) * 2006-12-21 2008-07-03 Plexxikon, Inc. Pyrrolo [2,3-b] pyridines as kinase modulators
PE20121126A1 (es) * 2006-12-21 2012-08-24 Plexxikon Inc Compuestos pirrolo [2,3-b] piridinas como moduladores de quinasa
PL2479267T3 (pl) 2006-12-21 2017-06-30 Basf Enzymes Llc Amylazy i glukoamylazy, kwasy nukleinowe kodujące te związki oraz sposoby wytwarzania tych związków oraz stosowania ich
JP2010514695A (ja) 2006-12-21 2010-05-06 プレキシコン，インコーポレーテッド キナーゼ調節のための化合物および方法およびそのための適応症
NZ598285A (en) 2007-01-30 2013-10-25 Syngenta Participations Ag Enzymes for the treatment of lignocellulosics, nucleic acids encoding them and methods for making and using them
US8916745B2 (en) 2007-04-27 2014-12-23 The Regents Of The University Of California Plant CO2 sensors, nucleic acids encoding them, and methods for making and using them
WO2009012283A1 (en) 2007-07-17 2009-01-22 Plexxikon Inc. Compounds and methods for kinase modulation, and indications therefor
US7507539B2 (en) * 2007-07-30 2009-03-24 Quest Diagnostics Investments Incorporated Substractive single label comparative hybridization
BRPI0817811A2 (pt) 2007-10-03 2014-10-14 Verenium Corp Xilanases, ácidos nucleicos que codificam as mesmas e métodos para fabricação e uso das mesmas.
CN104651381A (zh) 2008-01-03 2015-05-27 巴斯夫酶有限责任公司 转移酶和氧化还原酶、编码它们的核酸以及其制备和应用方法
US20090215050A1 (en) * 2008-02-22 2009-08-27 Robert Delmar Jenison Systems and methods for point-of-care amplification and detection of polynucleotides
US20110118125A1 (en) * 2008-05-03 2011-05-19 Tufts Medical Center, Inc. Neonatal salivary genomics
US8357503B2 (en) 2008-08-29 2013-01-22 Bunge Oils, Inc. Hydrolases, nucleic acids encoding them and methods for making and using them
DK2329259T3 (en) 2008-08-29 2016-05-09 Janssen Biotech Inc MARKERS AND PROCEDURES FOR ASSESSING AND TREATING ULCERATIVE COLITIS AND RELATED DISEASES USING A 20-GEN PANEL
US8198062B2 (en) 2008-08-29 2012-06-12 Dsm Ip Assets B.V. Hydrolases, nucleic acids encoding them and methods for making and using them
US8153391B2 (en) 2008-08-29 2012-04-10 Bunge Oils, Inc. Hydrolases, nucleic acids encoding them and methods for making and using them
US20120052066A1 (en) 2008-11-07 2012-03-01 Cesar Calderon Markers and methods for assessing and treating lupus patients susceptible to photoprovocation
CN102369296A (zh) 2009-03-09 2012-03-07 生物蛋白有限公司 Mirac蛋白
MY172424A (en) * 2009-04-03 2019-11-25 Hoffmann La Roche Propane- i-sulfonic acid {3- (4-chloro-phenyl)-1h-pyrrolo [2, 3-b] pyridine-3-carconyl] -2, 4-difluoro-phenyl} -amide compositions and uses thereof
NZ596459A (en) 2009-05-21 2013-11-29 Verenium Corp Phytases, nucleic acids encoding them and methods for making and using them
US8329724B2 (en) 2009-08-03 2012-12-11 Hoffmann-La Roche Inc. Process for the manufacture of pharmaceutically active compounds
US20110046009A1 (en) * 2009-08-24 2011-02-24 Perkinelmer Health Sciences, Inc. Methods for detecting dna methylation using encoded particles
US8222397B2 (en) * 2009-08-28 2012-07-17 Promega Corporation Methods of optimal purification of nucleic acids and kit for use in performing such methods
US8039613B2 (en) 2009-08-28 2011-10-18 Promega Corporation Methods of purifying a nucleic acid and formulation and kit for use in performing such methods
AU2010298000A1 (en) 2009-09-25 2012-04-05 Signature Genomics Laboratories Llc Multiplex (+/-) stranded arrays and assays for detecting chromosomal abnormalities associated with cancer and other diseases
UA111708C2 (uk) 2009-10-16 2016-06-10 Бандж Ойлз, Інк. Спосіб рафінування олії
UA109884C2 (uk) 2009-10-16 2015-10-26 Поліпептид, що має активність ферменту фосфатидилінозитол-специфічної фосфоліпази с, нуклеїнова кислота, що його кодує, та спосіб його виробництва і застосування
EP2491141A2 (en) 2009-10-19 2012-08-29 Stichting Het Nederlands Kanker Instituut Differentiation between brca2-associated tumours and sporadic tumours via array comparative genomic hybridization
EP2491140A1 (en) 2009-10-19 2012-08-29 Stichting Het Nederlands Kanker Instituut Predicting response to anti-cancer therapy via array comparative genomic hybridization
EP2491139A1 (en) 2009-10-19 2012-08-29 Stichting Het Nederlands Kanker Instituut Predicting benefit of anti-cancer therapy via array comparative genomic hybridization
CN106220623A (zh) 2009-11-06 2016-12-14 普莱希科公司 用于激酶调节的化合物和方法及其适应症
NL2004275C2 (en) * 2010-02-22 2011-08-23 Univ Leiden Raman spectrometry.
US20140018251A1 (en) 2010-09-20 2014-01-16 Stichting Het Nederlands Kanker Instituut Methods for Predicting Response to Anti-Cancer Therapy in Cancer Patients
US9096871B2 (en) 2010-10-06 2015-08-04 Bp Corporation North America Inc. Variant CBH I polypeptides with reduced product inhibition
US9624213B2 (en) 2011-02-07 2017-04-18 Plexxikon Inc. Compounds and methods for kinase modulation, and indications therefor
AR085279A1 (es) 2011-02-21 2013-09-18 Plexxikon Inc Formas solidas de {3-[5-(4-cloro-fenil)-1h-pirrolo[2,3-b]piridina-3-carbonil]-2,4-difluor-fenil}-amida del acido propano-1-sulfonico
JP6113151B2 (ja) 2011-05-17 2017-04-12 プレキシコン インコーポレーテッドＰｌｅｘｘｉｋｏｎ Ｉｎｃ． キナーゼ調節およびその適応症
WO2013067448A2 (en) 2011-11-03 2013-05-10 Vca Antech Inc. Compositions and methods to detect various infectious organisms
CA2859021A1 (en) 2011-12-19 2013-06-27 Valley Health System Methods and kits for detecting subjects at risk of having cancer
US9150570B2 (en) 2012-05-31 2015-10-06 Plexxikon Inc. Synthesis of heterocyclic compounds
US10670610B2 (en) 2012-06-15 2020-06-02 Wayne State University Biomarker test for prediction or early detection of preeclampsia and/or HELLP syndrome
HUE065221T2 (hu) 2012-06-15 2024-05-28 Genesis Theranostix Korlatolt Feleloessegue Tarsasag Biomarker teszt praeeclampsia elõrejelzésére vagy korai kimutatására
AU2013302861A1 (en) 2012-08-13 2015-03-05 The Rockefeller University Treatment and diagnosis of melanoma
WO2014071067A2 (en) 2012-10-31 2014-05-08 The Rockefeller University Treatment and diagnosis of colon cancer
US9267171B2 (en) 2013-02-28 2016-02-23 New York University DNA photolithography with cinnamate crosslinkers
ES2913205T3 (es) 2014-05-13 2022-06-01 Bioatla Inc Proteínas biológicas activas condicionalmente
US10745761B2 (en) 2014-06-02 2020-08-18 Valley Health System Method and systems for lung cancer diagnosis
DK4074735T3 (da) 2014-08-28 2025-07-14 Bioatla Inc Betinget aktive kimæriske antigenreceptorer til modificerede t-celler
US11111288B2 (en) 2014-08-28 2021-09-07 Bioatla, Inc. Conditionally active chimeric antigen receptors for modified t-cells
WO2016036916A1 (en) 2014-09-03 2016-03-10 Bioatla, Llc Discovering and producing conditionally active biologic proteins in the same eukaryotic cell production hosts
EP3262217A4 (en) 2015-02-24 2018-07-18 BioAtla LLC Conditionally active biological proteins
WO2016164641A1 (en) 2015-04-08 2016-10-13 Plexxikon Inc. Compounds and methods for kinase modulation, and indications therefor
US10829484B2 (en) 2015-07-28 2020-11-10 Plexxikon Inc. Compounds and methods for kinase modulation, and indications therefor
MX2018006856A (es) 2015-12-07 2018-08-01 Plexxikon Inc Compuestos y metodos para modulacion de la quinasa, e indicaciones de los mismos.
PL3455261T3 (pl) 2016-05-13 2022-12-12 Bioatla, Inc. Przeciwciała anty-ror2, fragmenty przeciwciał, ich immunokoniugaty oraz ich zastosowania
TW201815766A (zh) 2016-09-22 2018-05-01 美商普雷辛肯公司 用於ｉｄｏ及ｔｄｏ調節之化合物及方法以及其適應症
EP3558991A2 (en) 2016-12-23 2019-10-30 Plexxikon Inc. Compounds and methods for cdk8 modulation and indications therefor
WO2018226846A1 (en) 2017-06-07 2018-12-13 Plexxikon Inc. Compounds and methods for kinase modulation
CA3078981A1 (en) 2017-11-21 2019-05-31 Rgenix, Inc. Polymorphs and uses thereof
CA3087896A1 (en) 2018-01-09 2019-07-18 Mcmaster University Fluorosilinated liquid-infused surfaces with embedded biomolecules, methods of making and uses thereof
EP3594290A1 (en) 2018-07-13 2020-01-15 Haelixa GmbH Marked items and verification methods
US11884980B2 (en) 2018-11-28 2024-01-30 Bioscreening & Diagnostics Llc Method for detection of traumatic brain injury
TWI888447B (zh) 2019-12-13 2025-07-01 美商因思博納公司 金屬鹽及其用途
EP4368613A1 (fr) 2022-11-08 2024-05-15 aeChem Life Technologies Sàrl Dérivés de dicyanomethanides de phthalazinium
WO2025212475A2 (en) 2024-04-03 2025-10-09 Mars, Incorporated Compositions and methods to detect infectious organisms

Family Cites Families (39)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4231910A (en) * 1979-02-08 1980-11-04 Dow Corning Corporation Primer composition
CA1130228A (en) * 1980-05-21 1982-08-24 Chiang-Chang Liao Support matrix for amino-containing biologically active substances
CA1253129A (en) * 1984-02-09 1989-04-25 Thomas R. Jones Porous inorganic materials
US4957858A (en) * 1986-04-16 1990-09-18 The Salk Instute For Biological Studies Replicative RNA reporter systems
US4637687A (en) * 1984-06-14 1987-01-20 General Electric Company Cascaded, dual cell transflective liquid crystal display
US4818681A (en) * 1985-02-22 1989-04-04 Molecular Diagnostics, Inc. Fast and specific immobilization of nucleic acids to solid supports
US5641630A (en) * 1985-06-13 1997-06-24 Amgen Inc. Method and kit for performing nucleic acid hybridization assays
US4806631A (en) * 1985-09-30 1989-02-21 Miles Inc. Immobilization of nucleic acids on solvolyzed nylon supports
US4937188A (en) * 1986-04-15 1990-06-26 Northeastern University Enzyme activity amplification method for increasing assay sensitivity
US5190864A (en) * 1986-04-15 1993-03-02 Northeastern University Enzyme amplification by using free enzyme to release enzyme from an immobilized enzyme material
US4713116A (en) * 1987-01-02 1987-12-15 Ralston Purina Company Protein modified with a silanation reagent as an adhesive binder and process of producing
DE3880273T2 (de) * 1987-11-27 1993-07-29 Ecc Int Ltd Poroese anorganische materialien.
GB8803413D0 (en) * 1988-02-15 1988-03-16 Ecc Int Ltd Biological support
US5024933A (en) * 1988-05-10 1991-06-18 Enzo Biochem, Inc. Method and kit for sample adherence to test substrate
US5071909A (en) * 1989-07-26 1991-12-10 Millipore Corporation Immobilization of proteins and peptides on insoluble supports
US5252743A (en) * 1989-11-13 1993-10-12 Affymax Technologies N.V. Spatially-addressable immobilization of anti-ligands on surfaces
US5215882A (en) * 1989-11-30 1993-06-01 Ortho Diagnostic Systems, Inc. Method of immobilizing nucleic acid on a solid surface for use in nucleic acid hybridization assays
ES2116977T3 (es) * 1990-05-11 1998-08-01 Microprobe Corp Soportes solidos para ensayos de hibridacion de acidos nucleicos y metodos para inmovilizar oligonucleotidos de modo covalente.
DE4018778A1 (de) * 1990-06-12 1991-12-19 Braun Melsungen Ag Adsorptionsmaterial zur selektiven entfernung von ldl- oder/und vldl
IL102486A (en) * 1991-10-04 1997-11-20 Orgenics Ltd Method and apparatus for detection of nucleic acid sequences with a nucleic acid probe
US5632957A (en) * 1993-11-01 1997-05-27 Nanogen Molecular biological diagnostic systems including electrodes
US5412087A (en) * 1992-04-24 1995-05-02 Affymax Technologies N.V. Spatially-addressable immobilization of oligonucleotides and other biological polymers on surfaces
US5610287A (en) * 1993-12-06 1997-03-11 Molecular Tool, Inc. Method for immobilizing nucleic acid molecules
US6015880A (en) * 1994-03-16 2000-01-18 California Institute Of Technology Method and substrate for performing multiple sequential reactions on a matrix
US5807522A (en) * 1994-06-17 1998-09-15 The Board Of Trustees Of The Leland Stanford Junior University Methods for fabricating microarrays of biological samples
US5554744A (en) * 1994-09-23 1996-09-10 Hybridon, Inc. Method for loading solid supports for nucleic acid synthesis
US5688642A (en) * 1994-12-01 1997-11-18 The United States Of America As Represented By The Secretary Of The Navy Selective attachment of nucleic acid molecules to patterned self-assembled surfaces
US5807756A (en) * 1995-01-10 1998-09-15 At Point Bio Ceramic assembly for use in biological assays
US5601982A (en) * 1995-02-07 1997-02-11 Sargent; Jeannine P. Method and apparatus for determining the sequence of polynucleotides
US5959098A (en) * 1996-04-17 1999-09-28 Affymetrix, Inc. Substrate preparation process
US5630932A (en) * 1995-09-06 1997-05-20 Molecular Imaging Corporation Tip etching system and method for etching platinum-containing wire
US5658802A (en) * 1995-09-07 1997-08-19 Microfab Technologies, Inc. Method and apparatus for making miniaturized diagnostic arrays
US5851769A (en) * 1995-09-27 1998-12-22 The Regents Of The University Of California Quantitative DNA fiber mapping
US6156502A (en) * 1995-12-21 2000-12-05 Beattie; Kenneth Loren Arbitrary sequence oligonucleotide fingerprinting
US6426183B1 (en) * 1995-12-21 2002-07-30 Kenneth L. Beattie Oligonucleotide microarrays: direct covalent attachment to glass
US5567294A (en) * 1996-01-30 1996-10-22 Board Of Governors, University Of Alberta Multiple capillary biochemical analyzer with barrier member
AU734704B2 (en) * 1996-05-30 2001-06-21 Cellomics, Inc. Miniaturized cell array methods and apparatus for cell-based screening
US6048695A (en) * 1998-05-04 2000-04-11 Baylor College Of Medicine Chemically modified nucleic acids and methods for coupling nucleic acids to solid support
US6979728B2 (en) * 1998-05-04 2005-12-27 Baylor College Of Medicine Articles of manufacture and methods for array based analysis of biological molecules

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
BEATTIE W G, ET AL.: "HYBRIDIZATION OF DNA TARGETS TO GLASS-TETHERED OLIGONUCLEOTIDE PROBES", MOLECULAR BIOTECHNOLOGY, HUMANA PRESS, INC., US, vol. 04, 1 January 1995 (1995-01-01), US, pages 213 - 225, XP002919161, ISSN: 1073-6085, DOI: 10.1007/BF02779015 *
SHALON D, SMITH S J, BROWN P O: "A DNA MICROARRAY SYSTEM FOR ANALYZING COMPLEX DNA SAMPLES USING TWO-COLOR FLUORESCENT PROBE HYBRIDIZATION", GENOME RESEARCH, COLD SPRING HARBOR LABORATORY PRESS, vol. 06, 1 January 1996 (1996-01-01), pages 639 - 645, XP002919162, ISSN: 1088-9051 *

Cited By (24)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6858713B1 (en) 1998-05-04 2005-02-22 Baylor College Of Medicine Chemically modified biological molecules and methods for coupling biological molecules to solid support
AU2001263094B2 (en) * 1998-05-04 2008-07-10 Baylor College Of Medicine Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
US6979728B2 (en) 1998-05-04 2005-12-27 Baylor College Of Medicine Articles of manufacture and methods for array based analysis of biological molecules
EP1006363A3 (en) * 1998-12-01 2000-08-16 Hitachi Software Engineering Co., Ltd. Biochip and method for producing the same
EP1281967A3 (en) * 1998-12-01 2004-02-04 Hitachi Software Engineering Co., Ltd. Biochip and method for producing the same
DE19957827A1 (de) * 1999-11-25 2001-06-21 Epigenomics Ag Oligomer-Array mit PNA- und/oder DNA-Oligomeren auf einer Oberfläche
DE19957827C2 (de) * 1999-11-25 2003-06-12 Epigenomics Ag Verwendung eines Oligomer-Arrays mit PNA- und/oder DNA-Oligomeren auf einer Oberfläche
WO2001042501A1 (en) * 1999-12-09 2001-06-14 Karolinska Innovations Ab Method for immobilisation
EP1110967A1 (en) * 1999-12-21 2001-06-27 LION Bioscience AG Compound comprising a biomolecule moiety and an organo-silane moiety
WO2001046214A3 (en) * 1999-12-21 2001-12-27 Lion Bioscience Ag Compound comprising a nucleic acid moiety and an organo-silane moiety
WO2001046461A3 (en) * 1999-12-22 2002-05-23 Biochip Technologies Gmbh Modified nucleic acids and their use
JP2003521680A (ja) * 2000-01-11 2003-07-15 ナノゲン・レコグノミクス・ゲゼルシャフト・ミット・ベシュレンクテル・ハフツング 基体表面に結合させるための複数の結合部分を有する生体分子
JP2003520350A (ja) * 2000-01-20 2003-07-02 サントル・ナショナル・ドゥ・ラ・ルシェルシュ・シアンティフィク−シーエヌアールエス シラン化固体支持体上で核酸を合成および固定化するための方法
WO2002092615A3 (en) * 2001-05-10 2002-12-19 Baylor College Medicine Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
JP2004534959A (ja) * 2001-07-09 2004-11-18 サントル　ナショナル　ドゥ　ラ　ルシェルシュ　シアンティフィク 固形支持体の機能化方法、機能化された固形支持体、およびその使用
DE10139283A1 (de) * 2001-08-09 2003-03-13 Epigenomics Ag Verfahren und Nukleinsäuren zur Analyse von Colon-Krebs
US9671396B2 (en) * 2001-09-05 2017-06-06 Joon Won Park Solid substrate comprising array of dendrons and methods for using the same
EP1451318A4 (en) * 2001-10-12 2007-06-27 Perkinelmer Las Inc COMPOSITIONS OF NUCLEIC ACIDS AND ARRAYS AND METHOD FOR THEIR USE
US7217512B2 (en) 2002-05-09 2007-05-15 Corning Incorporated Reagent and method for attaching target molecules to a surface
US7195908B2 (en) 2002-10-31 2007-03-27 Corning Incorporated Supports treated with triamine for immobilizing biomolecules
WO2006072045A3 (en) * 2004-12-30 2006-08-24 Corning Inc Substrates having pendant epoxide groups for binding biomolecules and methods of making and using thereof
DE102008053270A1 (de) * 2008-10-27 2010-05-12 Medizinische Hochschule Hannover Vorrichtung und Verfahren zur Analyse von Zellen
FR3038734A1 (fr) * 2015-07-10 2017-01-13 Centre Nat De La Rech Scient (Cnrs) Nouveaux materiaux optiques fonctionnalises
WO2017009230A1 (fr) * 2015-07-10 2017-01-19 Universite De Montpellier Nouveaux materiaux optiques fonctionnalises

Also Published As

Publication number Publication date
CA2326684C (en) 2006-03-14
CA2326684A1 (en) 1999-11-11
EP1075544A4 (en) 2005-03-09
EP1075544B1 (en) 2011-11-30
ATE535615T1 (de) 2011-12-15
JP4477774B2 (ja) 2010-06-09
DE99920342T1 (de) 2005-01-20
AU770695B2 (en) 2004-02-26
US20050064494A1 (en) 2005-03-24
EP1075544A1 (en) 2001-02-14
JP2002513814A (ja) 2002-05-14
US6858713B1 (en) 2005-02-22
AU3786199A (en) 1999-11-23
US6048695A (en) 2000-04-11

Similar Documents

Publication Publication Date Title
EP1075544B1 (en) Chemically modified nucleic acids and methods for coupling nucleic acids to solid support
US6262216B1 (en) Functionalized silicon compounds and methods for their synthesis and use
US6387631B1 (en) Polymer coated surfaces for microarray applications
US20040106110A1 (en) Preparation of polynucleotide arrays
US6979728B2 (en) Articles of manufacture and methods for array based analysis of biological molecules
US6806361B1 (en) Methods of enhancing functional performance of nucleic acid arrays
JP3883539B2 (ja) エポキシ基を有する放射状ポリエチレングリコール誘導体を用いたハイドロゲルバイオチップの製造方法
US6426183B1 (en) Oligonucleotide microarrays: direct covalent attachment to glass
JP3398366B2 (ja) Ｄｎａ分析用マイクロアレイの製造方法
JP2003144172A (ja) メチル化検出用オリゴヌクレオチド固定化基板
EP1288664B1 (en) Methods for generating ligand arrays
Frydrych-Tomczak et al. Application of epoxy functional silanes in the preparation of DNA microarrays
EP4191638A1 (en) Surface linker of semiconductor chip, preparation method therefor and application thereof
US6989175B2 (en) Acyl fluoride activation of carboxysilyl-coated glass substrates
CA2446050A1 (en) Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
AU2001263094B2 (en) Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
AU2001263094A1 (en) Compositions and methods for array-based genomic nucleic acid analysis of biological molecules
US20030232361A1 (en) Nucleic acid array preparation using purified phosphoramidites
US20050119473A1 (en) Phosphite ester oxidation in nucleic acid array preparation

Legal Events

Date Code Title Description
AK Designated states

Kind code of ref document: A1

Designated state(s): AU CA JP

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE

121 Ep: the epo has been informed by wipo that ep was designated in this application
DFPE Request for preliminary examination filed prior to expiration of 19th month from priority date (pct application filed before 20040101)
WWE Wipo information: entry into national phase

Ref document number: 37861/99

Country of ref document: AU

ENP Entry into the national phase

Ref document number: 2326684

Country of ref document: CA

Ref country code: CA

Ref document number: 2326684

Kind code of ref document: A

Format of ref document f/p: F

ENP Entry into the national phase

Ref country code: JP

Ref document number: 2000 547274

Kind code of ref document: A

Format of ref document f/p: F

WWE Wipo information: entry into national phase

Ref document number: 1999920342

Country of ref document: EP

WWP Wipo information: published in national office

Ref document number: 1999920342

Country of ref document: EP

WWG Wipo information: grant in national office

Ref document number: 37861/99

Country of ref document: AU