US20070154946A1 - Massively parallel synthesis of biopolymeric arrays - Google Patents

Massively parallel synthesis of biopolymeric arrays Download PDF

Info

Publication number
US20070154946A1
US20070154946A1 US11/322,268 US32226805A US2007154946A1 US 20070154946 A1 US20070154946 A1 US 20070154946A1 US 32226805 A US32226805 A US 32226805A US 2007154946 A1 US2007154946 A1 US 2007154946A1
Authority
US
United States
Prior art keywords
peptide bond
forming
substrate
molecule
peptide
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.)
Abandoned
Application number
US11/322,268
Other languages
English (en)
Inventor
John Rajasekaran
Edelmira Cabezas
Jacqueline Fidanza
Gunjan Tiwari
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.)
Intel Corp
Original Assignee
Individual
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 Individual filed Critical Individual
Priority to US11/322,268 priority Critical patent/US20070154946A1/en
Priority to AT06845499T priority patent/ATE509940T1/de
Priority to PCT/US2006/047854 priority patent/WO2007078868A1/fr
Priority to EP06845499A priority patent/EP1966230B1/fr
Publication of US20070154946A1 publication Critical patent/US20070154946A1/en
Assigned to INTEL CORPORATION reassignment INTEL CORPORATION ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: RAJASEKARAN, JOHN J., CABEZAS, EDELMIRA, TIWARI, GUNJAN, FIDANZA, JACQUELINE A.
Priority to US12/133,260 priority patent/US20090258796A1/en
Priority to US12/133,219 priority patent/US20080242561A1/en
Priority to US12/133,190 priority patent/US9499578B2/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/04—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length on carriers
    • C07K1/047—Simultaneous synthesis of different peptide species; Peptide libraries

Definitions

  • the present invention relates generally to semiconductor lithographic technology, microarrays of bio-polymers, and synthetic organic chemistry.
  • Microarrays of oligonucleotides, peptides, proteins, and or oligosaccharides continue to gain importance as powerful tools for research and diagnostic applications in the biomedical sciences.
  • Oligonucleotide microarrays for example, can be used to monitor gene expression and genetic mutations in a massively parallel manner.
  • Proteinaceous microarrays provide the ability, for example, to characterize the molecular progression of disease, research cellular pathways, and perform high throughput screening in drug discovery applications.
  • Peptide-containing arrays can serve as molecular probes for a variety of biological events, such as for example, the arrays can serve as antigens for antibody-antigen systems, ligands for cell receptor-ligand systems, and substrates for enzyme-protein systems.
  • FIGS. 1A through 1F demonstrate a method for the controllable synthesis of polymers on a solid support involving semiconductor lithographic technology.
  • FIG. 2 provides chemical structure diagrams for exemplary molecules and functional groups.
  • FIG. 3 shows a method for derivatizing a SiO 2 surface and attaching a linker molecule to the derivatized surface.
  • FIG. 4 demonstrates general methods for solid phase peptide synthesis.
  • FIG. 5 demonstrates a general method for the synthesis of branched proteinaceous molecules on a solid surface that can be used to create arrays of molecules.
  • FIGS. 6A-6C provide exemplary bifurcation molecules.
  • FIG. 7 outlines an additional method for the synthesis of branched proteinaceous molecules on a solid surface that can be used to create arrays of biomolecules.
  • FIG. 8 diagrams the effect of the application of microwave energy to a substrate surface having attached polymers.
  • FIG. 9 diagrams the use of microwave energy in the photoresist method for peptide parallel synthesis.
  • An array is an intentionally-created collection of molecules housed on a solid support in which the identity or source of a group of molecules is known based on its location on the array.
  • the molecules housed on the array and within a feature of an array can be identical to or different from each other.
  • Embodiments of the present invention provide methods for the synthesis of polymers on a solid support. Polymer synthesis according to embodiments of the invention can be accomplished in a manner that provides controlled-density microarrays comprised of peptides, peptoids, peptidemimetics, branched peptides, and or other small bio-molecules.
  • Embodiments of the present invention provide arrays of biopolymers of known sequence and controllable molecular density. Additionally, the methods described in embodiments of the invention are highly scalable for array manufacture on a wafer or chip as are used in to fabricated devices in the semiconductor industry.
  • the features, regions, or sectors of an array in which the bio-polymers are located may have any convenient shape, for example, the features of the array may be circular, square, rectangular, elliptical, or wedge-shaped.
  • the region in which each distinct biomolecule is synthesized within a feature is smaller than about 1 mm 2 , or less than 0.5 mm 2 .
  • the features have an area less than about 10,000 ⁇ m 2 or less than 2.5 ⁇ m 2 .
  • multiple copies of a polymer will typically be located within any feature. The number of copies of a polymer can be in the thousands to the millions within a feature.
  • an array can have any number of features, and the number of features contained in an array may be selected to address such considerations as, for example, experimental objectives, information-gathering objectives, and cost effectiveness.
  • An array could be, for example, a 20 ⁇ 20 matrix having 400 regions, 64 ⁇ 32 matrix having 2,048 regions, or a 640 ⁇ 320 array having 204,800 regions.
  • the present invention is not limited to a particular size or configuration for the array.
  • FIGS. 1 A-F A method for synthesizing unbranched polymers within one or more selected region(s) of a solid support is shown in FIGS. 1 A-F.
  • the method includes attachment of a first amino acid, linker (or spacer) molecule, or other building block 2 to the surface of a substrate 1 .
  • a first building block 2 can be an amino acid that is attached to a substrate 1 that is comprised of amino-functionalized glass, through the formation of a peptide bond between the carboxylate of the amino acid and the amine group of the glass.
  • the terminal bond-forming site of the building block 2 is protected with a protecting group 3 .
  • the ⁇ -amino group of an amino acid can be protected with an N-protecting group 3 to prevent unwanted reactivity.
  • a side chain of the building block (for example, an R group of an amino acid) may also have a protecting group.
  • Suitable protecting groups include, for example, t-butoxycarbonyl (t-BOC) ( FIG. 2 , structure (II)), and fluorenylmethoxycarbonyl (FMOC) ( FIG. 2 , Structure (III), and 2-(4-biphenylyl)-2-oxycarbonyl.
  • embodiments of the present invention are not limited to the type of acid- or base-removable protective group or building block selected.
  • the photoresist layer can be created from a solution comprising a polymer, a photosensitizer, and a photo-active compound or molecule in a solvent.
  • the photoresist can be applied using any method known in the art of semiconductor manufacturing for the coating of a wafer with a photoresist layer, such as for example, the spin-coating method.
  • the photoresist-coated substrate is then baked to remove excess solvent from the photoresist and for film uniformity.
  • a photomask 5 is placed over photoresist layer 4 .
  • the photomask use may be practiced according to standard techniques and materials used in the semiconductor fabrication industry.
  • the photomask 5 may be a transparent pane, such as a quartz pane, having an emulsion or metal film on a surface creating the mask pattern. Suitable metals include chromium.
  • the pattern of the mask is chosen so that regions on the surface of the substrate can be selectively activated for polymer synthesis. Radiation, for example, ultra violet radiation (UV) or deep ultraviolet radiation (DUV), may then be directed through the photomask 5 onto the photoresist layer.
  • the photoresist 4 is exposed in those regions of the mask that are transparent to the impinging radiation.
  • the device used for creating a pattern in the photoresist can be a physical mask or any other source capable of projecting a pattern image, for example a micromirror.
  • the exposure of the photoresist 4 to radiation generates cleaving reagents (species that catalyze the removal of a protective group, for example) in the exposed portion of the photoresist layer 4 .
  • the generation of cleaving reagents in the photoresist may be the result of a number of processes.
  • the cleaving reagent may result from the direct radiation-induced decomposition of or chemical transformation of a photoactive cleavage reagent precursor compound.
  • generation of the cleaving reagent may occur through the absorption of light by a photosensitizer followed by reaction of the photosensitizer with the cleavage reagent precursor, energy transfer from the photosensitizer to the cleavage reagent precursor, or a combination of two or more different mechanisms.
  • the protecting groups 3 are cleaved from the molecules 2 under the exposed area(s) of the photoresist.
  • the molecules 2 located under the unexposed masked regions remain unreacted.
  • the cleaving process leading to the removal of the protecting groups 3 may, for example, be acid-catalyzed cleavage or base-catalyzed cleavage.
  • the chemistry of the process will depend on the type of protecting groups 3 and on the type of cleaving reagents that are generated in the photoresist upon radiation exposure. For example, if the protecting group 3 is t-BOC, acid cleavage can be used.
  • Acids may be generated in the photoresist, for example, through the exposure of sulfonium or halonium salts to radiation ( FIG. 2 , Structures (IV-VII) and (VIII-IX), respectively).
  • the protecting group is FMOC, for example, then base cleavage can be used. Cleavage can be accomplished through the reaction of a photogenerated amine or diamine through a decarboxylation process ( FIG. 5 ).
  • the rate of protecting group removal can be accelerated by heating the substrate after the exposure to radiation (post exposure bake).
  • the post exposure bake serves multiple purposes in photoresist processing. First, the elevated temperature of the bake drives diffusion of the photoproducts.
  • a small amount of diffusion can be useful in minimizing the effects of standing waves, periodic variations in exposure dose throughout the depth of the film that result from interference of incident and reflected radiation.
  • the other main purpose of the PEB is to drive the acid-catalyzed reaction. Chemical amplification is important because it allows a single photoproduct to cause many solubility-switching reactions, thus increasing the sensitivity of these photoresist systems.
  • the photoresist is removed.
  • the photoresist layer 4 may be removed using acetone or another similar suitable solvent.
  • the resulting surface-modified substrate is shown schematically in FIG. 1D .
  • FIG. 1D In this structure, there are three regions shown: two regions that have protected molecules and a region having deprotected molecules.
  • the deprotected molecules are available for further reaction, such as for example, a peptide-bond forming coupling reaction whereas the molecules that retain their protective groups are not available for further reaction.
  • FIG. 1E shows a structure resulting from the reaction of the deprotected surface-attached molecules.
  • a building block 6 has been added to molecule 2 .
  • Building block 6 may be the same or different from molecule 2 .
  • the building block 6 is protected with a protecting group to prevent unwanted reactions.
  • FIGS. 1 A-E may be repeated to form polymers on the substrate surface.
  • different polymers comprising building blocks 2 and 6 - 10 may be formed in regions upon the surface, as shown schematically in FIG. 2F .
  • the building blocks are amino acids
  • peptides having the same or different known sequences are formed in known regions on the surface of the substrate.
  • polymers containing from about 2 to about 50 mers (polymeric units) can be created.
  • peptides having a length of about 6 to about 20 amino acids are created.
  • capping reagents can be a reagent that prevents further reactivity at the site of polymer chain formation such as, for example, an acid anhydride without further reactive functionalities.
  • Capping groups cap deprotected functional groups by, for example, binding with the unreacted amino functions to form amides.
  • Capping agents suitable for use in an embodiment of the invention include: acetic anhydride, n-acetylimidizole, isopropenyl formate, fluorescamine, 3-nitrophthalic anhydride and 3 -sulfoproponic anhydride.
  • peptides are polymers of amino acids, amino acid mimics or derivatives, and/or unnatural amino acids.
  • the amino acids can be any amino acids, including ⁇ , ⁇ , or ⁇ -amino acids and modified amino acids.
  • the amino acids are ⁇ -amino acids, either the L-optical isomer or the D-optical isomer may be used.
  • FIG. 2 Structure (I), shows a general structural representation for an amino acid.
  • an amino acid contains an amine group, a carboxylic group, and an R group.
  • the R group can be a group found on a natural amino acid or a group that is similar in size to a natural amino acid R group.
  • unnatural amino acids for example, ⁇ -alanine, phenylglycine, homoarginine, aminobutyric acid, aminohexanoic acid, aminoisobutyric acid, butylglycine, citrulline, cyclohexylalanine, diaminopropionic acid, hydroxyproline, norleucine, norvaline, ornithine, penicillamine, pyroglutamic acid, sarcosine,and thienylalanine are also contemplated by the embodiments of the invention.
  • These and other natural and unnatural amino acids are available from, for example, EMD Biosciences, Inc., San Diego, Calif.
  • a peptide is a polymer in which the monomers are amino acids, a group of molecules which includes natural or unnatural amino acids, amino acid mimetics, and amino acid derivatives, which are generally joined together through amide (peptide) bonds.
  • a peptide can alternatively be referred to as a polypeptide.
  • Peptides contain two or more amino acid monomers, and often more than 50 amino acid monomers (building blocks).
  • a protein is a long polymer of amino acids linked via peptide bonds and which may be composed of one or more polypeptide chains. More specifically, the term protein refers to a molecule comprised of one or more polymers of amino acids. Proteins are essential for the structure, function, and regulation of the body's cells, tissues, and organs, and each protein has unique functions. Examples of proteins include some hormones, enzymes, and antibodies.
  • a protecting group is a group which is bound to a molecule and designed to block a reactive site in a molecule, but may be removed upon exposure to an activator or a deprotecting reagent.
  • Deprotecting reagents include, for example, acids and bases.
  • Protecting groups can be bound to a monomer, a polymer, a linker molecule or a monomer, or polymer, or a linker molecule attached to a solid support to protect a reactive functionality on the monomer, polymer, or linker molecule.
  • Protective groups that may be used in accordance with an embodiment of the invention include all acid and base labile protecting groups.
  • peptide amine groups are preferably protected by t-butoxycarbonyl (t-BOC or BOC) (shown in FIG. 1 , Structure (II)) or benzyloxycarbonyl (CBZ), both of which are acid labile, or by 9-fluorenylmethoxycarbonyl (FMOC) (shown in FIG. 1 , Structure (III)), which is base labile.
  • t-BOC or BOC t-butoxycarbonyl
  • BOC benzyloxycarbonyl
  • FMOC 9-fluorenylmethoxycarbonyl
  • Additional protecting groups that may be used in accordance with embodiments of the invention include acid labile groups for protecting amino moieties: tert-amyloxycarbonyl, adamantyloxycarbonyl, 1-methylcyclobutyloxycarbonyl, 2-(p-biphenyl)propyl(2)oxycarbonyl, 2-(p-phenylazophenylyl)propyl(2)oxycarbonyl, .alpha.,.alpha.-dimethyl-3,5-dimethyloxybenzyloxy-carbonyl, 2-phenylpropyl(2)oxycarbonyl, 4-methyloxybenzyloxycarbonyl, furfuryloxycarbonyl, triphenylmethyl (trityl), p-toluenesulfenylaminocarbonyl, dimethylphosphinothioyl, diphenylphosphinothioyl, 2-benzoyl-1-methylvinyl, o-nitrophenylsulfenyl, and 1-na
  • a linker molecule typically is a molecule inserted into the growing polymer that does not necessarily convey functionality to the resulting peptide, such as molecular recognition functionality, but instead elongates the distance between the substrate surface and the peptide functionality to enhance the exposure of the peptide functionality on the surface of the substrate.
  • a linker should be about 4 to about 40 atoms long to provide exposure.
  • the linker molecules may be, for example, aryl acetylene, ethylene glycol oligomers containing 2-10 monomer units (PEGs), diamines, diacids, amino acids, among others, and combinations thereof. Examples of diamines include ethylene diamine and diamino propane.
  • the linkers may be the same molecule type as that being synthesized (i.e., nascent polymers), such as polypeptides and polymers of amino acid derivatives such as for example, amino hexanoic acids.
  • Solid support, support, and substrate refer to a material or group of materials having a rigid or semi-rigid surface or surfaces. In some aspects, at least one surface of the solid support will be substantially flat, although in some aspects it may be desirable to physically separate synthesis regions for different molecules with, for example, wells, raised regions, pins, etched trenches, or the like. In certain embodiments, the solid support may be porous.
  • Substrate materials useful in embodiments of the present invention include, for example, silicon, bio-compatible polymers such as, for example poly(methyl methacrylate) (PMMA) and polydimethylsiloxane (PDMS), glass, SiO 2 (such as, for example, a thermal oxide silicon wafer such as that used by the semiconductor industry), quartz, silicon nitride, functionalized glass, gold, platinum, and aluminum.
  • Functionalized surfaces include for example, amino-functionalized glass, carboxy functionalized glass, and hydroxy functionalized glass.
  • a substrate may optionally be coated with one or more layers to provide a surface for molecular attachment or functionalization, increased or decreased reactivity, binding detection, or other specialized application.
  • Substrate materials and or layer(s) may be porous or non-porous.
  • a substrate may be comprised of porous silicon.
  • the substrate may be a silicon wafer or chip such as those used in the semiconductor device fabrication industry. In the case of a wafer or chip, a plurality of arrays may be synthesized on the wafer.
  • Photoresist formulations useful in the present invention include a polymer, a solvent, and a radiation-activated cleaving reagent.
  • Useful polymers include, for example, poly(methyl methacrylate) (PMMA), poly-(methyl isopropenyl ketone) (PMPIK), poly-(butene-1-sulfone) (PBS), poly-(trifluoroethyl chloroacrylate) (TFECA), copolymer-( ⁇ -cyano ethyl acrylate- ⁇ -amido ethyl acrylate (COP), and poly-(2-methyl pentene-1-sulfone).
  • Useful solvents include, for example, propylene glycol methyl ether acetate (PGMEA), ethyl lactate, and ethoxyethyl acetate.
  • the solvent used in fabricating the photoresist may be selected depending on the particular polymer, photosensitizer, and photo-active compound that are selected.
  • the polymer used in the photoresist is PMMA
  • the photosensitizer is isopropyl-thioxanthenone
  • the photoactive compound is diphenyliodonium chloride
  • PGMEA or ethyl lactate may be used as the solvent.
  • the mass concentration of the polymer may between about 5% and about 50%, the mass concentration of a photosensitizer may be up to about 20%, the mass concentration of the photo-active compound may be between about 1% and 10%, the balance comprising a suitable solvent.
  • the substrate typically is heated to form the photoresist layer. Any method known in the art of semiconductor fabrication may be used to for depositing the photoresist solution. For example, the spin coating method may be used in which the substrate is spun typically at speeds between about 1,000 and about 5,000 revolutions per minute for about 30 to about 60 seconds.
  • the resulting wet photoresist layer has a thickness ranging between about 0.1 ⁇ m to about 2.5 ⁇ m.
  • Catalysts for protective group removal include acids and bases.
  • acids can be generated photochemically from sulfonium salts ( FIG. 2 , Structures IV-VII), halonium salts ( FIG. 2 , Structures VIII-IX), and polonium salts ( FIG. 2 , Structures X-XI).
  • Sulfonium ions are positive ions, R 3 S + , where R is, for example, a hydrogen or alkyl group, such as methyl, phenyl, or other aryl group.
  • Trimethyl sulfonium iodide and triaryl sulfonium hexafluroantimonatate are shown in FIG. 2 , Structures VII and VI, respectively.
  • halonium ions are bivalent halogens, R 2 X + , where R is a hydrogen or an alkyl group, such as methyl, phenyl, or other aryl group, and X is a halogen atom.
  • the halonium ion may be linear or cyclic.
  • Polonium salt refers to a halonium salt where the halogen is iodine, the compound R 2 I + Y ⁇ , where Y is an anion, for example, a nitrate, chloride, or bromide.
  • FIG. 2 shows diphenyliodonium chloride and diphenyliodonium nitrate (Structure X and XI, respectively). See also, Frechet, J. M. J., Ito, H., Willson, C. G., Proc. Microcircuit Eng., 260, (1982); Shirai, M., Tsunooka, M., Prog. Polym. Sci., 21:1, (1996); Frechet, J. M.
  • Photogenerated bases include amines and diamines having photolabile protecting groups. See for example, Shirai, M., Tsunooka, M., Prog. Polym. Sci., 21:1, (1996); Comeron, J. F., Frechet, J. M. J., J. Org. Chem., 55:5919, (1990); Comeron, J. F., Frechet, J. M. J., J. Am. Chem. Soc., 113:4303, (1991); and Arimitsu, K. and Ichimura, K., J. Mat. Chem,. 14:336, (2004).
  • the photoresists useful in the present invention may also include a photosensistizer.
  • a photosensitizer absorbs light and interacts with the cleavage reagent precursor, through one or more mechanisms, including, reaction with the cleavage reagent precursor and energy transfer from the photosensitizer to the cleavage reagent precursor.
  • Useful photosensitizers include, for example, benzophenone ( FIG. 2 , Structure XII) and other similar diphenyl ketones, thioxanthenone ( FIG. 2 , Structure XIII), isopropyl-thioxanthenone, anthraquinone, fluorenone, acetophenone, and perylene.
  • FIG. 3 provides a method for derivatization of a SiO 2 surface and linking of polymeric molecules to the surface.
  • the SiO 2 surface is silanated by reacting it with aminopropyltriethoxy silane (APTES).
  • APTES aminopropyltriethoxy silane
  • the resulting surface presents an amine functional group for further reaction, such as peptide bond formation.
  • Modulation of the density of polymers on the surface can be attained by silanation.
  • density can be modulated by mixing a functionalizable silane for example, APTES, with a non-functional silane (a silane with no non silyl functional group), for example, propyltrialkoxy silane.
  • the derivatized surface can then be reacted with a linker.
  • the linker is a polyethylene glycol molecule having an amine group protected with BOC at one terminus and a peptide-bond forming group (a carboxyl group) at the second terminus.
  • This coupling reaction can be accomplished in a solution of carboxyl group activators: 1-hydroxybenzotriazole (HOBt) and diisopropylcarbodiimide (DIC) in N-methyl pyrrolidone (NMP).
  • HOBt 1-hydroxybenzotriazole
  • DIC diisopropylcarbodiimide
  • NMP N-methyl pyrrolidone
  • FIG. 4 shows a general scheme for solid-phase peptide synthesis.
  • a substrate surface is provided having a first amino acid attached to the surface.
  • a second amino acid having an amino protecting group is coupled to the first amino acid.
  • the second amino acid is N-protected with a BOC protecting group.
  • the coupling reaction is performed in a solution of 1-hydroxybenzotriazole (HOBt) and diisopropylcarbodiimide (DIC) in N-methyl pyrrolidone (NMP). Unreacted amine groups are capped using an acetic anhydride (Ac 2 0 ) solution in dimethylformamide (DMF).
  • the substrate surface is then coated with a photoresist.
  • the photoresist is comprised of PMMA polymer, TASSbF 6 (photoactivated acid generator), and PGMEA (as a solvent).
  • TFA represents trifluoroacetic acid, the acidic cleaving reagent typically used for bulk solid-phase peptide synthesis.
  • yields for a peptide synthesis process according to the current invention were similar to yields for bulk solid-phase synthesis procedures using TFA as a protecting group removal catalyst.
  • UV radiation an acid is produced in the photoresist and the N-protecting group is removed from the attached peptide in the region of UV exposure.
  • FIG. 5 diagrams a method for using semiconductor photoresist acid and base cleavage mechanisms and orthogonal protection chemistries to create a branch point in a growing solid-phase polymer chain.
  • a region of a substrate surface presents an attached amino acid building block containing an acid-cleavable protecting group that prevents the formation of a peptide bond.
  • the amino acid building block may be attached to the surface through a linker molecule that spaces the amino acid from the surface and or that functions as a peptide or peptidemimetic.
  • the substrate surface is coated with an acid-generating photoresist and at least one region of the substrate surface is exposed to radiation to induce the formation of the photo-generated acid in the photoresist.
  • a mask is used to selectively block the radiation from areas of the substrate surface and thereby cause deprotection of only molecules located in selected regions of the substrate surface.
  • the substrate surface is subjected to a post-exposure bake.
  • the photoresist is then stripped from the substrate surface leaving a section(s) of the substrate having deprotected sites available for the formation of peptide bonds.
  • a bifurcation group is coupled to the deprotected growing chain.
  • the bifurcation group contains both acid and base cleavable protecting groups.
  • the branch points of the bifurcation group can be selectively deprotected by selecting either an acid or a base-generating photoresist. In the scheme shown in FIG.
  • a base-generating photoresist is selected and the base-cleavable protecting group is removed from the bifurcation group upon irradiation.
  • the surface is baked and the resist removed.
  • An amino acid having a base-removable protecting group is coupled to the bifurcation group at the point in which the base-removable group was removed.
  • the acid labile protecting group may be removed in a similar manner using a photoresist containing a photogenerated acid as described herein.
  • An amino acid having an acid-removable protecting group is then coupled to the bifurcation group. Repetitions of these acid or base deprotection-coupling events allow a branched polymer having a desired sequence to be synthesized.
  • the polymers formed from the branch-point may be the same or different lengths.
  • optionally polymers having one or more branch points may be created.
  • An array may be created having both branched and unbranched polymers in regions upon its surface.
  • a bifurcation molecule is a molecule that when it is attached to a growing polymer provides more than one location for further polymer growth or extension.
  • a bifurcation molecule is a polymeric building block that creates a branch point in a polymer.
  • FIGS. 6 A-C provide several exemplary bifurcation molecules: FIG. 6A shows Trt-Dpr(Fmoc)-OH (Dpr is diaminopropionic acid); FIG. 6B shows Fmoc-Dab(Mtt)-OH (“Dab” is diaminobutanoic acid, “Mtt” is 4-methyltrityl, an acid-removable protecting group); FIG.
  • bifurcation molecules include Fmoc-Dpr(Boc)-OH, Fmoc-Dab(Boc)-OH, or any of the previous amino acids having the same protecting group at the amino and the side chain functionalities (to allow growth of two peptide chains having the same sequence), such as for example, Fmoc-Dpr(Fmoc)-OH or Trt-Dpr(Trt)-OH.
  • Fmoc-Dpr(Fmoc)-OH or Trt-Dpr(Trt)-OH are available from EMD Biosciences Nova Biochem, La Jolla, Calif.; Bachem, Torrance, Calif.; and Chem-Impex International.
  • FIG. 7 diagrams a further method by which branch points can be created in a polymer using orthogonal protection strategies and solid-phase bio-polymer synthesis.
  • direct photo-cleavage lithography is applied to the synthesis of a branched peptide array.
  • a region of a substrate surface presents an attached amino acid building block containing a photo-cleavable protecting group that prevents the formation of a peptide bond.
  • the amino acid building block may be attached to the surface through a linker molecule that spaces the amino acid from the surface and or that functions as a peptide or peptidemimetic.
  • the photo-cleavable protecting group is removable at a first wavelength, ⁇ 1 .
  • a selected area(s) of the substrate surface are exposed to light of wavelength ⁇ 1 and the ⁇ 1 photo-removable protecting group is removed.
  • An amino acid building block having a ⁇ 1 photo-removable protecting group is coupled to the deprotected site of the bifurcation group. Repetitions of these photo-deprotection-coupling events allow a branched polymer having a desired sequence to be synthesized.
  • the polymers formed from the branch-point may be the same or different lengths. Additionally, optionally polymers having one or more branch points may be created.
  • An array may be created having both branched and unbranched polymers in regions upon its surface.
  • wavelength controlled orthogonal protecting groups are described in “Wavelength-Controlled Orthogonal Photolysis of Protecting Groups,” Blanc, A. and Bochte, C., J. Org. Chem., 67:5567 (2002). Additional examples of photo-cleavable protecting groups can be found in: Photogenerated Reagents in Biochemistry and Molecular Biology , Bayley, H., Laboratory Techniques in Biochemistry and Molecular Biology, 12, Elsevier (2002); and Clark, M. A. and Romoff, T. T., Synthesis of peptides and peptidomimetics, Methods of Organic Chemistry , E22a:276 (2004). The most common photocleavable groups are 2-nitrobenzyl-derived carbamates, esters, and ethers.
  • Examples include Z(2-NO2), Nvoc, Ndmoc, Dnboc, Menpoc, Ddz.
  • Other examples include tosyl, phenacyl-based esters, and the 5-bromo-7-nitro-indolinyl group.
  • Many amino acids with photocleavable protecting groups are available from Chem-Impex International, Wood Dale, Ill.
  • methods are provided to synthesize biopolymers on an array wherein the synthesis is assisted by microwave radiation.
  • Organic synthesis reactions that are assisted by microwave radiation generally demonstrate faster rates and improved fidelity.
  • Synthesis of biopolymers, particularly parallel synthesis methodologies can encounter difficulties, depending on the species being synthesized, from intermolecular aggregation, hydrogen bonded secondary structures (such as, for example, beta-sheet formation in peptides), steric hindrance from bulky side chain protecting groups, and hence premature termination of the sequence.
  • FIG. 8 a schematic is presented showing the microwave heating of polymers on a substrate surface. In this example, the heating of the polymers causes them to unwind.
  • microwave energy directly activates any molecule with a dipole moment and allows for local heating at a molecular level.
  • intermolecular and intramolecular aggregation, secondary structure formation, and steric hindrance can be overcome with microwave energy.
  • reaction times for deprotection and coupling reactions can be reduced and fidelity of lithographic techniques can be enhanced.
  • FIG. 9 shows the use of microwave energy to assist the synthesis of biopolymers on a solid support.
  • microwave energy can be used to facilitate the synthesis of branched and unbranched biopolymers, such as for example, peptides, peptidemimetics, and polynucleotides, by photoresist methods and methods employing photo-removable protecting groups, as described herein and as known in the art.
  • polynucleotide includes oligonucleotides, peptide nucleic acids (PNAs), and cDNA.
  • a substrate surface presents attached biopolymer building blocks having a protecting group that prevents polymer formation. The substrate is coated with a photoresist.
  • Synthesizers generally supply microwave power of about 25 to 50 W and a temperature range of 20 to 70° C. Additionally, microwave energy can be employed in the manufacture of arrays on silicon chips. In this case a microwave stepper and a microwave synthesizer can be used to manufacture a plurality of arrays in a similar manner as it is employed in the semiconductor fabrication industry.
  • a glass substrate was silanated using a solution of 3% APTES (aminopropyl triethoxy silane) in 95% ethanol. The surface of the substrate was then washed and annealed at about 100° C. for about 1 hour. The substrate was then treated with a 1:1 solution of DIEA (diisopropyl ethyl amine) in DMF (dimethylformamide).
  • APTES aminopropyl triethoxy silane
  • a spacer molecule was then coupled to the surface using a solution of 0.25 M solution of O-(N-Boc-2-aminoethyl)-O′-(N-diglycolyl-2-aminoethyl)hexaethyleneglycol, 0.25 M HOBt, and 0.25 M DIC (diisopropylcarbodiimide) in NMP (N-methyl pyrrolidone)and gentle agitation over the surface of the substrate in a sealed container for about 30 min. The solution was then discarded and the surface replenished with fresh solution. After coupling was complete, the surface was washed with NMP and then acetone. Unreacted surface amine groups were capped by treatment with 1:1 acetic anhydride in DMF solution (a 50% acetic anhydride solution in DMF) for about 30 minutes. The surface was then washed.
  • a photoresist was prepared by mixing about 10% by mass of PMMA, 20% by mass of triarylsulfonium hexafluoroantimonate in PGMEA solvent and spin coating the mixture over the amino acid derivatized glass surface for about 60 seconds at 2,000 rpm.
  • the photoresist may also optionally contain thioxanthenone, a photosensitizer.
  • the photoresist layer was baked at about 85° C. for about 90 seconds. The resulting photoresist layer had a thickness of about 2 ⁇ m.
  • Acid was generated in the photoresist layer by irradiation of the surface of the substrate with 2-3 J of 365 nm UV light through a mask. The reaction was accelerated by a post exposure bake at about 65° C. for about 60 seconds. After the photogenerated acid deprotection was achieved, the surface of the substrate was rinsed with acetone to strip the photoresist from the surface and the surface was dried. The surface was neutralized by treatment in 25% DIEA/DMF for about 5-10 minutes and then washed in DMF.
  • a second amino acid (Boc-Leu-OH) was coupled to the surface of the substrate using a 0.25 M solution of N- ⁇ -Boc-Leu, HOBt, and DIC as above. Subsequent rounds of coupling and deprotection were accomplished by repeating the above procedures to generate peptides of a desired length.
  • a hexamer peptide SDLYKL segement of human tumor suppressor protein p53
  • PEG polyethylene glycol
  • the peptide array was incubated for 1 hour with 5 ⁇ g/ml monoclonal antibody known to specifically recognize the SDLHKL epitope of human p53 protein. A second incubation was performed with fluorescein-labeled rabbit antibody raised against mouse antibody at a 1:100 dilution in phosphate buffered saline with 0.05% Tween 20. A fluorescent checker board pattern was detected on fluorescence scanning of the array suggesting specific interaction of antibody with the wildtype sequence.
  • Photoresist formulations may include a sensitizer in addition to the photogenerated acid catalyst to generate the acid deprotection catalysts.
  • the amount of PMMA in the resist in these exemplary formulations may vary between about 3% and about 50%.
  • Useful photoresists may be made using diaryliodonium salts (DAI) and photosensitizers.
  • the mass ration between DAI and photosensitizer may be between about 1:10 and 1:1.
  • DAI diaryliodonium salts
  • Photosensitizers may be between about 1:10 and 1:1.
  • (tolylcumyl)polonium tetrakis (pentafluorophenyl)borate with isopropyl-9H-thioxanthen-9-one may be formulated in a 1:10 or 1:1 (or a ratio there between) in PMMA and PGMEA to form final concentrations of between about 0.5% to 10% by mass DAI.
  • the formulation selected may be spun coated on the substrate surface and baked.
  • the radiation exposure dose may be between about 0.02 J and about 10 J.
  • Post exposure baking may be conducted for about 30 to 60 seconds at about 40° C. to about 85° C.

Landscapes

  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Analytical Chemistry (AREA)
  • Peptides Or Proteins (AREA)
US11/322,268 2005-12-29 2005-12-29 Massively parallel synthesis of biopolymeric arrays Abandoned US20070154946A1 (en)

Priority Applications (7)

Application Number Priority Date Filing Date Title
US11/322,268 US20070154946A1 (en) 2005-12-29 2005-12-29 Massively parallel synthesis of biopolymeric arrays
AT06845499T ATE509940T1 (de) 2005-12-29 2006-12-15 Massiv parallele synthese biopolymerer arrays
PCT/US2006/047854 WO2007078868A1 (fr) 2005-12-29 2006-12-15 Synthèse massivement parallèle de matrices biopolymères
EP06845499A EP1966230B1 (fr) 2005-12-29 2006-12-15 Synthese massivement parallele de matrices biopolymeres
US12/133,260 US20090258796A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,219 US20080242561A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,190 US9499578B2 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
US11/322,268 US20070154946A1 (en) 2005-12-29 2005-12-29 Massively parallel synthesis of biopolymeric arrays

Related Child Applications (3)

Application Number Title Priority Date Filing Date
US12/133,190 Division US9499578B2 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,219 Division US20080242561A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,260 Division US20090258796A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays

Publications (1)

Publication Number Publication Date
US20070154946A1 true US20070154946A1 (en) 2007-07-05

Family

ID=38008220

Family Applications (4)

Application Number Title Priority Date Filing Date
US11/322,268 Abandoned US20070154946A1 (en) 2005-12-29 2005-12-29 Massively parallel synthesis of biopolymeric arrays
US12/133,190 Expired - Fee Related US9499578B2 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,260 Abandoned US20090258796A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,219 Abandoned US20080242561A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays

Family Applications After (3)

Application Number Title Priority Date Filing Date
US12/133,190 Expired - Fee Related US9499578B2 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,260 Abandoned US20090258796A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays
US12/133,219 Abandoned US20080242561A1 (en) 2005-12-29 2008-06-04 Massively parallel synthesis of biopolymeric arrays

Country Status (4)

Country Link
US (4) US20070154946A1 (fr)
EP (1) EP1966230B1 (fr)
AT (1) ATE509940T1 (fr)
WO (1) WO2007078868A1 (fr)

Cited By (23)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20070275411A1 (en) * 2006-05-25 2007-11-29 Mcgall Glenn H Silane mixtures
US20080161202A1 (en) * 2006-12-29 2008-07-03 Edelmira Cabezas Novel strategy for selective regulation of background surface property in microarray fabrication and method to eliminated self quenching in micro arrays
US20080160446A1 (en) * 2006-12-28 2008-07-03 Meagley Robert P Photoresists including amino acid polymers as photoimageable species
US20080157786A1 (en) * 2006-12-28 2008-07-03 Gordon Holt Quality control methods for the manufacture of polymer arrays
US20080318808A1 (en) * 2005-12-29 2008-12-25 Rajasekaran John J Massively parallel synthesis of biopolymeric arrays
US20090093381A1 (en) * 2006-12-28 2009-04-09 Wei Wang Solid phase electrochemical synthesis with controlled product cleavage
US20090176664A1 (en) * 2007-06-01 2009-07-09 Keting Chu High density peptide arrays containing kinase or phosphatase substrates
US20100240555A1 (en) * 2006-09-29 2010-09-23 Narayan Sundararajan Method for high throughput, high volume manufacturing of biomolecule micro arrays
US20100240544A1 (en) * 2006-09-29 2010-09-23 Liu David J Aptamer biochip for multiplexed detection of biomolecules
US20100248975A1 (en) * 2006-12-29 2010-09-30 Gunjan Tiwari Fluorogenic peptide substrate arrays for highly multiplexed, real-time monitoring of kinase activities
US20110003716A1 (en) * 2002-06-20 2011-01-06 Affymetrix, Inc. Antireflective coatings for high-resolution photolithographic synthesis of dna array
WO2014078606A2 (fr) 2012-11-14 2014-05-22 Vibrant Holdings, Llc Substrats, systèmes et procédés pour la synthèse de réseau et l'analyse biomoléculaire
WO2014127328A2 (fr) 2013-02-15 2014-08-21 Vibrant Holdings, Llc. Procédés et compositions pour détection amplifiée d'une électrochimiluminescence
WO2015038919A1 (fr) * 2013-09-15 2015-03-19 Stc. Unm Méthode de dépôt de couches atomiques
WO2015127409A1 (fr) 2014-02-21 2015-08-27 Vibrant Holdings, Llc Photo-activation sélective d'acides aminés pour un couplage peptidique en une seule étape
EP2812344A4 (fr) * 2012-02-07 2015-10-28 Vibrant Holdings Llc Substrats, réseaux de peptides et procédés
US9417236B2 (en) 2012-09-28 2016-08-16 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
WO2016145434A1 (fr) 2015-03-12 2016-09-15 Vibrant Holdings, Llc Puces à polypeptides et procédés de fixation de polypeptides à une puce
CN106117453A (zh) * 2016-06-08 2016-11-16 广州高通生物技术有限公司 一种高分子母液及3d生物芯片的制作方法
US10006909B2 (en) 2012-09-28 2018-06-26 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
US10286376B2 (en) 2012-11-14 2019-05-14 Vibrant Holdings, Llc Substrates, systems, and methods for array synthesis and biomolecular analysis
WO2019217704A1 (fr) 2018-05-09 2019-11-14 Vibrant Holdings, Llc Procédés de synthèse d'un réseau de polynucléotides à l'aide d'agents photactivés
US11168365B2 (en) 2017-05-26 2021-11-09 Vibrant Holdings, Llc Photoactive compounds and methods for biomolecule detection and sequencing

Families Citing this family (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB201000879D0 (en) * 2010-01-20 2010-03-10 Univ Reading The Improved hydrogel synthesis
US10427125B2 (en) 2011-05-09 2019-10-01 Arizona Board Of Regents On Behalf Of Arizona State University Methods for performing patterned chemistry
WO2014062981A1 (fr) 2012-10-17 2014-04-24 Arizona Board Of Regents, A Body Corporate Of The State Of Arizona, Acting For And On Behalf Of Arizona State University Formation de motifs in situ par voie chimique
US10758886B2 (en) 2015-09-14 2020-09-01 Arizona Board Of Regents On Behalf Of Arizona State University Conditioned surfaces for in situ molecular array synthesis
KR102632268B1 (ko) * 2016-01-11 2024-01-31 삼성전자주식회사 포토 레지스트 조성물 및 이를 이용한 반도체 장치의 제조 방법
US11420941B2 (en) 2017-06-29 2022-08-23 Cowper Sciences Inc. Methods and systems for mask alignment in manufacturing process of arrays
US11517873B2 (en) 2019-10-29 2022-12-06 International Business Machines Corporation Controlled chemical synthesis using polymer substrates and nanofluidic separation systems

Citations (17)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3767398A (en) * 1971-10-26 1973-10-23 C Morgan Solid photoresist comprising a polyene and a polythiol
US5143854A (en) * 1989-06-07 1992-09-01 Affymax Technologies N.V. Large scale photolithographic solid phase synthesis of polypeptides and receptor binding screening thereof
US6083697A (en) * 1996-11-14 2000-07-04 Affymetrix, Inc. Chemical amplification for the synthesis of patterned arrays
US6121048A (en) * 1994-10-18 2000-09-19 Zaffaroni; Alejandro C. Method of conducting a plurality of reactions
US6239273B1 (en) * 1995-02-27 2001-05-29 Affymetrix, Inc. Printing molecular library arrays
US6359125B1 (en) * 1999-06-07 2002-03-19 Samsung Electronics Co., Ltd. Process for preparing peptide nucleic acid probe using polymeric photoacid generator
US6379895B1 (en) * 1989-06-07 2002-04-30 Affymetrix, Inc. Photolithographic and other means for manufacturing arrays
US6406844B1 (en) * 1989-06-07 2002-06-18 Affymetrix, Inc. Very large scale immobilized polymer synthesis
US6444111B1 (en) * 1996-07-05 2002-09-03 Combimatrix Corporation Electrochemical solid phase synthesis of polymers
US6506558B1 (en) * 1990-03-07 2003-01-14 Affymetrix Inc. Very large scale immobilized polymer synthesis
US6515039B1 (en) * 1998-08-28 2003-02-04 Poly-An Gmbh Method for the parallel and combinatory synthesis of compounds bound to a continuous polymeric solid phase supporting material
US20030082633A1 (en) * 2000-10-03 2003-05-01 Martin Mark T. Directed microwave chemistry
US6566495B1 (en) * 1989-06-07 2003-05-20 Affymetrix, Inc. Very large scale immobilized polymer synthesis
US20040198637A1 (en) * 2002-12-22 2004-10-07 The Scripps Research Institute Protein arrays
US20050026202A1 (en) * 2000-09-27 2005-02-03 Edman Carl F. Electronic systems and component devices for macroscopic and microscopic molecular biological reaction, analyses, and diagnostics
US6919181B2 (en) * 2002-03-25 2005-07-19 Agilent Technologies, Inc. Methods for generating ligand arrays
US7452679B2 (en) * 2003-12-05 2008-11-18 Northwestern University Branched peptide amphiphiles, related epitope compounds and self assembled structures thereof

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US6919211B1 (en) * 1989-06-07 2005-07-19 Affymetrix, Inc. Polypeptide arrays
US5959098A (en) * 1996-04-17 1999-09-28 Affymetrix, Inc. Substrate preparation process
EP1051148B1 (fr) * 1998-12-08 2004-06-30 Firmenich S.A. Composition parfumante transparente
DE19922941A1 (de) * 1999-05-14 2000-11-30 Epigenomics Gmbh Vorrichtung und Verfahren zur photolithographischen Belichtung von biologischen Stoffen
DE19925372A1 (de) * 1999-06-02 2000-12-07 Bosch Gmbh Robert Verfahren zum elektronischen Überwachen und Steuern eines Prozesses zum Verstellen beweglicher Teile
US20030044798A1 (en) 2001-08-31 2003-03-06 Lefkowitz Steven M. Methods for generating ligand arrays via deposition of ligands onto olefin displaying substrates, and arrays produced thereby
WO2003094031A1 (fr) * 2002-05-03 2003-11-13 Netbotz, Inc. Procede et appareil de collecte et d'affichage d'informations d'un dispositif reseau
US7202358B2 (en) 2002-07-25 2007-04-10 Agilent Technologies, Inc. Methods for producing ligand arrays
US20070034513A1 (en) 2005-03-25 2007-02-15 Combimatrix Corporation Electrochemical deblocking solution for electrochemical oligomer synthesis on an electrode array
US20070154946A1 (en) * 2005-12-29 2007-07-05 Rajasekaran John J Massively parallel synthesis of biopolymeric arrays
US7622295B2 (en) * 2006-12-19 2009-11-24 Edelmira Cabezas Molecular microarrays and helical peptides

Patent Citations (21)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3767398A (en) * 1971-10-26 1973-10-23 C Morgan Solid photoresist comprising a polyene and a polythiol
US5143854A (en) * 1989-06-07 1992-09-01 Affymax Technologies N.V. Large scale photolithographic solid phase synthesis of polypeptides and receptor binding screening thereof
US5405783A (en) * 1989-06-07 1995-04-11 Affymax Technologies N.V. Large scale photolithographic solid phase synthesis of an array of polymers
US6630308B2 (en) * 1989-06-07 2003-10-07 Affymetrix, Inc. Methods of synthesizing a plurality of different polymers on a surface of a substrate
US6566495B1 (en) * 1989-06-07 2003-05-20 Affymetrix, Inc. Very large scale immobilized polymer synthesis
US6379895B1 (en) * 1989-06-07 2002-04-30 Affymetrix, Inc. Photolithographic and other means for manufacturing arrays
US6406844B1 (en) * 1989-06-07 2002-06-18 Affymetrix, Inc. Very large scale immobilized polymer synthesis
US6416952B1 (en) * 1989-06-07 2002-07-09 Affymetrix, Inc. Photolithographic and other means for manufacturing arrays
US6506558B1 (en) * 1990-03-07 2003-01-14 Affymetrix Inc. Very large scale immobilized polymer synthesis
US6121048A (en) * 1994-10-18 2000-09-19 Zaffaroni; Alejandro C. Method of conducting a plurality of reactions
US6239273B1 (en) * 1995-02-27 2001-05-29 Affymetrix, Inc. Printing molecular library arrays
US6444111B1 (en) * 1996-07-05 2002-09-03 Combimatrix Corporation Electrochemical solid phase synthesis of polymers
US6083697A (en) * 1996-11-14 2000-07-04 Affymetrix, Inc. Chemical amplification for the synthesis of patterned arrays
US6770436B1 (en) * 1996-11-14 2004-08-03 Affymetrix, Inc. Chemical amplification for the synthesis of patterned arrays
US6515039B1 (en) * 1998-08-28 2003-02-04 Poly-An Gmbh Method for the parallel and combinatory synthesis of compounds bound to a continuous polymeric solid phase supporting material
US6359125B1 (en) * 1999-06-07 2002-03-19 Samsung Electronics Co., Ltd. Process for preparing peptide nucleic acid probe using polymeric photoacid generator
US20050026202A1 (en) * 2000-09-27 2005-02-03 Edman Carl F. Electronic systems and component devices for macroscopic and microscopic molecular biological reaction, analyses, and diagnostics
US20030082633A1 (en) * 2000-10-03 2003-05-01 Martin Mark T. Directed microwave chemistry
US6919181B2 (en) * 2002-03-25 2005-07-19 Agilent Technologies, Inc. Methods for generating ligand arrays
US20040198637A1 (en) * 2002-12-22 2004-10-07 The Scripps Research Institute Protein arrays
US7452679B2 (en) * 2003-12-05 2008-11-18 Northwestern University Branched peptide amphiphiles, related epitope compounds and self assembled structures thereof

Cited By (67)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20110003716A1 (en) * 2002-06-20 2011-01-06 Affymetrix, Inc. Antireflective coatings for high-resolution photolithographic synthesis of dna array
US10213762B2 (en) 2002-06-20 2019-02-26 Affymetrix, Inc. Silane mixtures
US9556360B2 (en) 2002-06-20 2017-01-31 Affymetrix, Inc. Silane mixtures
US8859196B2 (en) 2002-06-20 2014-10-14 Affymetrix, Inc. Antireflective coatings for high-resolution photolithographic synthesis of DNA arrays
US8114584B2 (en) 2002-06-20 2012-02-14 Affymetrix, Inc. Antireflective coatings for high-resolution photolithographic synthesis of DNA array
US8105821B2 (en) 2002-06-20 2012-01-31 Affymetrix, Inc. Silane mixtures
US20080318808A1 (en) * 2005-12-29 2008-12-25 Rajasekaran John J Massively parallel synthesis of biopolymeric arrays
US9499578B2 (en) 2005-12-29 2016-11-22 Intel Corporation Massively parallel synthesis of biopolymeric arrays
US20090258796A1 (en) * 2005-12-29 2009-10-15 Rajasekaran John J Massively parallel synthesis of biopolymeric arrays
US20070275411A1 (en) * 2006-05-25 2007-11-29 Mcgall Glenn H Silane mixtures
US9096953B2 (en) 2006-09-29 2015-08-04 Intel Corporation Method for high throughput, high volume manufacturing of biomolecule micro arrays
US20100240544A1 (en) * 2006-09-29 2010-09-23 Liu David J Aptamer biochip for multiplexed detection of biomolecules
US20100240555A1 (en) * 2006-09-29 2010-09-23 Narayan Sundararajan Method for high throughput, high volume manufacturing of biomolecule micro arrays
US8603803B2 (en) 2006-12-28 2013-12-10 Intel Corporation Solid phase electrochemical synthesis with controlled product cleavage
US20110224092A1 (en) * 2006-12-28 2011-09-15 Intel Corporation Method and apparatus for combined electrochemical synthesis and detection of analytes
US8338097B2 (en) 2006-12-28 2012-12-25 Intel Corporation Method and apparatus for combined electrochemical synthesis and detection of analytes
US8486631B2 (en) 2006-12-28 2013-07-16 Intel Corporation Quality control methods for the manufacture of polymer arrays
US8614086B2 (en) 2006-12-28 2013-12-24 Intel Corporation Quality control methods for the manufacture of polymer arrays
US8647821B2 (en) 2006-12-28 2014-02-11 Intel Corporation Method and apparatus for combined electrochemical synthesis and detection of analytes
US20080160446A1 (en) * 2006-12-28 2008-07-03 Meagley Robert P Photoresists including amino acid polymers as photoimageable species
US20080157786A1 (en) * 2006-12-28 2008-07-03 Gordon Holt Quality control methods for the manufacture of polymer arrays
US20090093381A1 (en) * 2006-12-28 2009-04-09 Wei Wang Solid phase electrochemical synthesis with controlled product cleavage
US20080161202A1 (en) * 2006-12-29 2008-07-03 Edelmira Cabezas Novel strategy for selective regulation of background surface property in microarray fabrication and method to eliminated self quenching in micro arrays
US20100248975A1 (en) * 2006-12-29 2010-09-30 Gunjan Tiwari Fluorogenic peptide substrate arrays for highly multiplexed, real-time monitoring of kinase activities
US20090176664A1 (en) * 2007-06-01 2009-07-09 Keting Chu High density peptide arrays containing kinase or phosphatase substrates
US12454771B2 (en) 2012-02-07 2025-10-28 Vibrant Holdings Llc Substrates, peptide arrays, and methods
EP2812344A4 (fr) * 2012-02-07 2015-10-28 Vibrant Holdings Llc Substrats, réseaux de peptides et procédés
US9216399B2 (en) 2012-02-07 2015-12-22 Vibrant Holdings, Llc Substrates, peptide arrays, and methods
US10486129B2 (en) 2012-02-07 2019-11-26 Vibrant Holdings, Llc Substrates, peptide arrays, and methods
US11565231B2 (en) 2012-02-07 2023-01-31 Vibrant Holdings, Llc Substrates, peptide arrays, and methods
US11674956B2 (en) 2012-09-28 2023-06-13 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
US10746732B2 (en) 2012-09-28 2020-08-18 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
US9417236B2 (en) 2012-09-28 2016-08-16 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
EP3690442A1 (fr) 2012-09-28 2020-08-05 Vibrant Holdings, LLC Procédés, systèmes et réseaux pour analyse biomoléculaire
US10175234B2 (en) 2012-09-28 2019-01-08 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
EP3974564A1 (fr) 2012-09-28 2022-03-30 Vibrant Holdings, LLC Procédés, systèmes et réseaux pour analyse biomoléculaire
US11815512B2 (en) 2012-09-28 2023-11-14 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
US10006909B2 (en) 2012-09-28 2018-06-26 Vibrant Holdings, Llc Methods, systems, and arrays for biomolecular analysis
WO2014078606A3 (fr) * 2012-11-14 2014-08-07 Vibrant Holdings, Llc Substrats, systèmes et procédés pour la synthèse de réseau et l'analyse biomoléculaire
JP2018119136A (ja) * 2012-11-14 2018-08-02 ヴィブラント ホールディングス リミテッド ライアビリティ カンパニー アレイ合成および生体分子解析のための、支持体、システム、および方法
US10286376B2 (en) 2012-11-14 2019-05-14 Vibrant Holdings, Llc Substrates, systems, and methods for array synthesis and biomolecular analysis
JP7514280B2 (ja) 2012-11-14 2024-07-10 ヴィブラント ホールディングス リミテッド ライアビリティ カンパニー アレイ合成および生体分子解析のための、支持体、システム、および方法
JP2024124458A (ja) * 2012-11-14 2024-09-12 ヴィブラント ホールディングス リミテッド ライアビリティ カンパニー アレイ合成および生体分子解析のための、支持体、システム、および方法
EP3617219A1 (fr) 2012-11-14 2020-03-04 Vibrant Holdings, LLC Procédés pour synthèse de réseau
US12251674B2 (en) 2012-11-14 2025-03-18 Vibrant Holdings, Llc Substrates, systems, and methods for array synthesis and biomolecular analysis
JP2016503447A (ja) * 2012-11-14 2016-02-04 ヴィブラント ホールディングス リミテッド ライアビリティ カンパニー アレイ合成および生体分子解析のための、支持体、システム、および方法
EP4556467A2 (fr) 2012-11-14 2025-05-21 Vibrant Holdings, LLC Procédés de synthèse de réseau
US10799845B2 (en) 2012-11-14 2020-10-13 Vibrant Holdings, Llc Substrates, systems, and methods for array synthesis and biomolecular analysis
EP4556467A3 (fr) * 2012-11-14 2025-08-13 Vibrant Holdings, LLC Procédés de synthèse de réseau
WO2014078606A2 (fr) 2012-11-14 2014-05-22 Vibrant Holdings, Llc Substrats, systèmes et procédés pour la synthèse de réseau et l'analyse biomoléculaire
JP2023001123A (ja) * 2012-11-14 2023-01-04 ヴィブラント ホールディングス リミテッド ライアビリティ カンパニー アレイ合成および生体分子解析のための、支持体、システム、および方法
US10816553B2 (en) 2013-02-15 2020-10-27 Vibrant Holdings, Llc Methods and compositions for amplified electrochemiluminescence detection
US12546781B2 (en) 2013-02-15 2026-02-10 Vibrant Holding, LLC Methods and compositions for amplified electrochemiluminescence detection assays
EP4030167A1 (fr) 2013-02-15 2022-07-20 Vibrant Holdings, LLC Procédés et compositions de détection de l'électrochimioluminescence amplifiée
EP3702782A2 (fr) 2013-02-15 2020-09-02 Vibrant Holdings, LLC Procédés et compositions de détection de l'électrochimioluminescence amplifiée
WO2014127328A2 (fr) 2013-02-15 2014-08-21 Vibrant Holdings, Llc. Procédés et compositions pour détection amplifiée d'une électrochimiluminescence
US9605344B2 (en) 2013-09-15 2017-03-28 Stc.Unm Method for atomic layer deposition
WO2015038919A1 (fr) * 2013-09-15 2015-03-19 Stc. Unm Méthode de dépôt de couches atomiques
WO2015127409A1 (fr) 2014-02-21 2015-08-27 Vibrant Holdings, Llc Photo-activation sélective d'acides aminés pour un couplage peptidique en une seule étape
EP3805207A1 (fr) 2014-02-21 2021-04-14 Vibrant Holdings, LLC Formulation pour la photo-activation sélective d'acides aminés pour un couplage peptidique en une seule étape
EP4170348A1 (fr) 2015-03-12 2023-04-26 Vibrant Holdings, LLC Réseaux de polypeptides et procédés de fixation de polypeptides à un réseau
WO2016145434A1 (fr) 2015-03-12 2016-09-15 Vibrant Holdings, Llc Puces à polypeptides et procédés de fixation de polypeptides à une puce
CN106117453A (zh) * 2016-06-08 2016-11-16 广州高通生物技术有限公司 一种高分子母液及3d生物芯片的制作方法
US12152279B2 (en) 2017-05-26 2024-11-26 Vibrant Holdings, Llc Photoactive compounds and methods for biomolecule detection and sequencing
US11168365B2 (en) 2017-05-26 2021-11-09 Vibrant Holdings, Llc Photoactive compounds and methods for biomolecule detection and sequencing
US12319712B2 (en) 2018-05-09 2025-06-03 Vibrant Holdings, Llc Methods of synthesizing a polynucleotide array using photoactivated agents
WO2019217704A1 (fr) 2018-05-09 2019-11-14 Vibrant Holdings, Llc Procédés de synthèse d'un réseau de polynucléotides à l'aide d'agents photactivés

Also Published As

Publication number Publication date
US20080318808A1 (en) 2008-12-25
EP1966230B1 (fr) 2011-05-18
ATE509940T1 (de) 2011-06-15
US9499578B2 (en) 2016-11-22
WO2007078868A1 (fr) 2007-07-12
US20080242561A1 (en) 2008-10-02
US20090258796A1 (en) 2009-10-15
EP1966230A1 (fr) 2008-09-10

Similar Documents

Publication Publication Date Title
US9499578B2 (en) Massively parallel synthesis of biopolymeric arrays
US20100035771A1 (en) Molecular microarrays and helical peptides
US20070122842A1 (en) Massively parallel synthesis of proteinaceous biomolecules
US20070122841A1 (en) Massively parallel synthesis of proteinaceous biomolecules
US20260071353A1 (en) Substrates, peptide arrays, and methods
US10577391B2 (en) Selective photoactivation of amino acids for single step peptide coupling
WO2008097370A2 (fr) Procédé de fabrication à haut débit et grand volume de microréseaux de biomolécules

Legal Events

Date Code Title Description
AS Assignment

Owner name: INTEL CORPORATION, CALIFORNIA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:RAJASEKARAN, JOHN J.;CABEZAS, EDELMIRA;FIDANZA, JACQUELINE A.;AND OTHERS;REEL/FRAME:019560/0986;SIGNING DATES FROM 20060329 TO 20060508

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION