WO2008023907A1 - Puce à protéines en nanoréseau et procédé de préparation de celle-ci - Google Patents

Puce à protéines en nanoréseau et procédé de préparation de celle-ci Download PDF

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Publication number
WO2008023907A1
WO2008023907A1 PCT/KR2007/003960 KR2007003960W WO2008023907A1 WO 2008023907 A1 WO2008023907 A1 WO 2008023907A1 KR 2007003960 W KR2007003960 W KR 2007003960W WO 2008023907 A1 WO2008023907 A1 WO 2008023907A1
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Prior art keywords
protein
nanoarray
substrate
target protein
chip
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PCT/KR2007/003960
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English (en)
Inventor
Hyung-Il Jung
Kyung-Hee Kim
Jung-Dong Kim
Sung-Kyu Kim
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Industry-Academic Cooperation Foundation, Yonsei University
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Publication of WO2008023907A1 publication Critical patent/WO2008023907A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54373—Apparatus specially adapted for solid-phase testing involving physiochemical end-point determination, e.g. wave-guides, FETS, gratings
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J19/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J19/0046—Sequential or parallel reactions, e.g. for the synthesis of polypeptides or polynucleotides; Apparatus and devices for combinatorial chemistry or for making molecular arrays
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00351—Means for dispensing and evacuation of reagents
    • B01J2219/00387—Applications using probes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00277—Apparatus
    • B01J2219/00497—Features relating to the solid phase supports
    • B01J2219/00527—Sheets
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00599—Solution-phase processes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00583—Features relative to the processes being carried out
    • B01J2219/00603—Making arrays on substantially continuous surfaces
    • B01J2219/00605—Making arrays on substantially continuous surfaces the compounds being directly bound or immobilised to solid supports
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01J—CHEMICAL OR PHYSICAL PROCESSES, e.g. CATALYSIS OR COLLOID CHEMISTRY; THEIR RELEVANT APPARATUS
    • B01J2219/00—Chemical, physical or physico-chemical processes in general; Their relevant apparatus
    • B01J2219/00274—Sequential or parallel reactions; Apparatus and devices for combinatorial chemistry or for making arrays; Chemical library technology
    • B01J2219/00718—Type of compounds synthesised
    • B01J2219/0072—Organic compounds
    • B01J2219/00725—Peptides

Definitions

  • the present invention relates to a protein chip. More particularly, the present invention relates to a protein nanoarray chip and a fabrication method thereof, in which protein patterns are formed at the nanometer scale using a pin microarray or inkjet method without purification of a target protein to be immobilized.
  • DNA chips have been fabricated and used to obtain information about genes associated with specific diseases, but had limitations in identifying the properties and activities of proteins, which are associated directly with diseases and play an important role in cells. For this reason, many researchers have developed protein chips either for analyzing proteomics at the protein level or for the diagnosis of specific diseases and the discovery of biomarkers. The protein chips are expected to become more useful tools than the DNA chips in studies on the expression and function of proteins and studies on protein- protein interactions, as well as the development of new drugs, the diagnosis of diseases, and food and environmental monitoring.
  • the term "protein chip” is defined as a system in which several tens to several hundreds of kinds of different proteins or ligands, which bind to specific proteins, are immobilized on a solid substrate made of, for example, a glass, metal or plastic material, such that the presence, function or role of biomolecules , which react specifically with the immobilized proteins or ligands, is rapidly analyzed using fluorescence, SPR (surface plasma resonance), mass spectrometry or the like.
  • the protein chip has technical characteristics similar to those of a DNA microarray chip in which several tens to several hundreds of genes are immobilized on a glass or plastic substrate, such that the expression or mutation thereof is identified by quantitatively analyzing the interactions between the genes.
  • Protein patterns can be formed according to the prior techniques, which use various methods, including photolithography, microcontact printing and spot arraying.
  • protein spots are formed by injecting target proteins to be immobilized, into microtubes in pins by capillary action, and then bringing the pins into direct contact with a substrate, or are formed directly on a substrate using a piezoelectric spray process.
  • proteins that are immobilized would necessarily be limited to the micrometer scale (65-200 ⁇ m) .
  • very precise positioning should be possible with a robot arm, which used in microarraying, prints one spot and then brings another sample, it can print the next spot while precisely maintaining an interval of several tens or several hundreds of nanometers .
  • Mirkin et al provided a protein nanoarray chip by forming nanometer-size (i.e., less than 100 nm) patterns on a substrate with an atomic force microscope (AFM) probe using DPN (dip-pen nanolithography) other than the prior pin array or inkjet methods ("Protein Nanoarrays Generated by Dip-Pen Nanolithography", Science, Vol. 295, March 2002, 1702-1705) .
  • AFM atomic force microscope
  • a protein nanoarray chip is fabricated by forming nanospots with a substance, to which a target protein to be immobilized can specifically bind, for example, l ⁇ -mercaptohexadecanoic acid (MHA) , and then allowing the target protein to bind selectively to the nanospot region.
  • MHA l ⁇ -mercaptohexadecanoic acid
  • this method that uses a high affinity between the carboxyl terminal end of MHA and the target protein has shortcomings in that it cannot control the orientation of the target protein, such that the active site of the protein can be immobilized on a substrate, and thus the activity of the protein can be lost, and in addition, the application thereof is limited only to proteins having a high affinity for the carboxyl terminal end of MHA. Also, in the method, different target proteins cannot be independently bound to a plurality of different MHA nanoarrays, and thus there is a limitation in fabricating a nano-sized protein chip immobilized with different target proteins .
  • a protein nanoarray chip is fabricated using a specific reaction between Ni 2+ ions and histidine, by coating the entire surface of a substrate with Ni, and then forming nanoarrays on the surface with histidine-conjugated target proteins by dip- pen nanolithography using AFM probes.
  • the activity of the target protein binding to the nanoarray can be maintained, because an affinity tag, such as histidine, which can bind to the target protein without reducing the activity of the target protein, is used.
  • an affinity tag such as histidine
  • the use of a plurality of AFM probes enables a nanoprotein chip to be fabricated with different histidine- conjugated target proteins.
  • the number of different target proteins formed on the substrate is limited to the number of AFM probes, because the AFM probes cannot be coated with other target proteins after washing, unlike with the pin microarray or inkjet methods.
  • the methods for fabricating the protein chips according to the prior technique have the following problems: i) the methods essentially comprise purifying proteins; ii) it is difficult to form protein nanopatterns; and iii) in the case of the DPN method that uses AFM probes, although nanosized protein patterns are formed, the number of target proteins to be immobilized on a substrate is limited depending on the number of AFM probes, making it impossible to fabricate a highly integrated protein chip, and in addition, it is difficult to prevent proteins from binding to sites other than the target sites.
  • the present inventors have fabricated a protein nanoarray chip in a way different from the prior technique, by making a nanoarray of a probe capable of binding to a target protein, on a substrate, spotting the target protein on the substrate at the same location as the probe nanoarray using a microarrayer, and washing out the region of the substrate, in which the target protein spots did not bind to the probe nanoarray, and the present inventors have confirmed the activity of the protein, immobilized on the substrate by the probe, thereby completing the present invention.
  • the present invention provides an epochal method, which can completely solve the problems occurring in the prior technique and can make nanoarrays of a plurality of different target proteins, and it can realize protein nanoarrays using the prior automated microarrayer and enable proteins to be applied in chip fabrication processes without purification.
  • the present invention provides solutions to the fundamental problems that several research groups for developing protein nanoarray chips have not solved for several years .
  • Another object of the present invention is to provide a protein nanoarray chip fabricated according to said method.
  • the present invention provides a highly integrated protein nanoarray chip, which is fabricated by making a nanoarray of a probe, for example, NTA/Ni 2+ , on a glass or plastic substrate, spotting a target protein, for example, histidine-conjugated target protein, on the probe nanoarray pattern (e.g., NTA/Ni 2+ pattern), using a pin array or inkjet method, and then washing the resulting substrate.
  • a target protein for example, histidine-conjugated target protein
  • the substrate can be made of one selected from among various materials, including glass, plastics, metals, organic thin films and other insulators.
  • glutathione, immunoglobulin (IgG), maltose or the like can be used for the probe on the substrate in the present invention, because glutathione S-transferase (GST) , protein-A and a maltose binding protein (MBP) bind specifically to substrate glutathione, immunoglobulin and maltose, respectively, and are affinity tags which are typically used in protein purification.
  • DPN printing in the present invention commercially available contact AFM probes are used to provide DPN patterns having a diameter or width of nanometers, for example, less than 100 nm, preferably 50-80 ran, and more preferably 20 nm.
  • the protein nanoarray chip according to the present invention is fabricated using a method comprising the steps of: i) making a nanoarray of a probe for binding to a target protein, on a substrate, to form nanopatterns having a size of 20-100 nm; ii) passivating a non-nanoarray portion on the substrate with a substance for preventing the non-specific binding of a target protein; iii) applying to the substrate either a solution containing a conjugate of the target protein with an affinity tag, which binds specifically to the probe, or an affinity tag- target protein conjugate purified from the solution, to immobilize the protein on the substrate; and iv) washing the substrate, the fabricated protein chip having a protein nanoarray with a spot density of 1,000-10,000 spots/cuf, and preferably 2,500-6,400 spots/cm 2 .
  • a protein can be nanoarrayed on a substrate, and thus a protein chip having increased sensitivity can be realized at the nanometer scale. Also, because the target protein to be immobilized can be used without purification, the protein nanoarray chip of the present invention can be used as a very useful means in, for example, proteomics, disease diagnosis and immunoassay. [Description of Drawings]
  • FIG. 1 shows a process of making a nanoarray using DPN according to the present invention.
  • FIG. 2 shows a process of making a nanoarray using DPN, and the sequential binding of MPTMS [3- (mercaptopropyl) - trimethoxysilane] , maleimido) -C3-NTA, Ni ions, and histidine-protein, on a glass surface.
  • FIG. 3 is a photograph showing an NTA/Ni 2+ nanoarray on a glass substrate.
  • FIG. 4 is a photograph showing that histidine-EGFP (enhanced green fluorescence protein) binds selectively to an NTA/Ni 2+ nanoarray.
  • FIG. 5 shows a process of making a plurality of NTA/Ni 2+ nanoarrays using a plurality of AFM probes .
  • FIG. 6 shows a process of fabricating a protein chip by spotting a protein on the NTA/Ni 2+ nanoarray using a pin array or inkjet printing method.
  • FIG. 7 shows a process of fabricating a protein chip by washing a protein chip spotted with different proteins.
  • FIG. 8 shows that histidine-protein binds to the NTA/Ni 2+ nanoarray without purification, after it is expressed in a bacterial strain.
  • FIG. 9 shows that histidine-conjugated HRP (horseradish peroxidase) has activity on a glass substrate nanopatterned with NTA/Ni 2+ .
  • FIG. 10 shows that a fabricated NTA/Ni 2+ nanoarray can be reused. Specifically, FIG. 10 shows the comparison of activity between the histidine-fluorescence protein, rebound to the NTA/Ni 2+ nanoarray (FIG. 10 (c) ) after washing with imidazole (FIG. 10 (b) ) , and the histidine- fluorescence protein before washing (FIG. 10 (a)).
  • FIG. 11 shows that a fluorescence protein as a target protein is applied using an mRNA-target protein fusion molecule without purification and binds to a surface having an oligonucleotide nanoarray pattern thereon.
  • FIG. 12 illustrates the results of SDS-PAGE detection of a reaction solution expressed in vitro, which show the expression of a plurality of proteins in addition to the target protein.
  • FIG. 1 shows a process of making a nanoarray using DPN according to the present invention.
  • FIG. 2 shows the sequential binding of MPTMS [3- (mercaptopropyl ) - trimethoxysilane) ] , maleimido-C3-NTA, Ni ions, and histidine-protein, on a glass surface.
  • MPTMS [3- (mercaptopropyl ) - trimethoxysilane)
  • maleimido-C3-NTA maleimido-C3-NTA
  • Ni ions ions
  • histidine-protein histidine-protein
  • the maleimido-C3-NTA is then nanoarrayed on the substrate, and an NTA nanoarray is formed on the substrate surface by a covalent bond between the maleimide and the -SH group, formed only in the nanoarray region.
  • the NTA nanoarray is treated with nickel (II) chloride hexahydrate, Ni 2+ ions are chelated to the NTA moiety, so that the surface is substituted with an NTA/Ni 2+ nanoarray.
  • FIG. 3 shows the NTA nanoarray on the glass substrate. As can be seen in FIG.
  • the size of nanoarray spots can be controlled depending to the concentration of the surfactant (e.g., Tween 20 (Sigma Aldrich) ) , the contact time between the probe and the substrate, etc.
  • FIG. 4 shows that histidine-EGFP (enhanced green fluorescence protein) binds selectively to the NTA/Ni 2+ nanoarray.
  • FIG. 5 shows a process of fabricating a plurality of NTA/Ni 2+ nanoarrays using a plurality of AFM probes.
  • FIG. 6 shows a process of fabricating a protein chip by spotting a protein on the NTA/Ni 2+ nanoarray using the prior pin array or inkjet printing method. In FIG.
  • the protein that is spotted using the pin array method is histidine-protein, and is spotted on the NTA/Ni 2+ nanoarrays fabricated as shown in FIG. 5.
  • FIG. 6 shows this process, in which histidine- protein 1 is spotted at a specific location on a substrate 1, and the same protein is spotted at a specific location on a substrate 2.
  • the pin is washed, histidine-protein 2 is loaded into the pin, and then the histidine-protein 2 is spotted at a position different from the histidine-protein using the pin array method.
  • FIG. 7 shows a process of fabricating a protein nanoarray chip by washing a protein chip spotted with different proteins.
  • FIG. 8 shows that histidine-protein used in the fabrication of a protein nanoarray chip binds to the NTA/Ni 2+ nanoarray without purification, after it is expressed in a bacterial strain.
  • FIG. 9 shows that the target protein is nanoarrayed using various biomolecules, including a histidine-conjugated protein, an enzyme and DNA, in addition to the fluorescence protein. As can be seen in FIG.
  • FIG. 10 shows that a fabricated NTA/Ni 2+ nanoarray can be re-used.
  • FIG. 11 shows a process in which a fluorescence protein as a target protein binds specifically to an oligonucleotide nanoarray using an mRNA-target protein fusion molecule without purification.
  • FIG. 12 shows that a target protein in a reaction solution, expressed in vitro, binds specifically to oligonucleotide nanopatterns .
  • histidine-protein refers to a histidine-conjugated protein.
  • target protein refers to a protein to be immobilized on a substrate.
  • affinity tag refers to a moiety, which is conjugated to the target protein and binds specifically to a probe nanoarrayed on a substrate.
  • Each of a glass substrate and an AFM probe was cleaned with a piranha solution (70% sulfuric acid and 30% hydrogen peroxide) and UV light, and then placed in a closed container, in which they were coated with 5 ⁇ l of MPTMS at 120 ° C for 30 minutes.
  • the AFM probe substituted with a thiol group (-SH) was coated with maleimido-C3-NTA ink for 10 minutes, and then dried with nitrogen gas, thus preparing an AFM probe coated with maleimido-C3-NTA.
  • the glass substrate substituted with a thiol group (-SH) was nanopatterned with maleimido-C3-NTA using DPN (Bio-AFM, DI product) technology.
  • DPN Bio-AFM, DI product
  • FIG. 3 shows the maleimido-C3-NTA nanopatterns .
  • the non-patterned region on the nanoarrayed glass substrate was passivated with PEG-maleimide .
  • the prepared patterns were allowed to react with 50 mM of nickel (II) chloride hexahydrate, thus fabricating NTA/Ni 2+ nanoarray patterns on the glass substrate.
  • the fabricated NTA/Ni 2+ nanoarray patterns were allowed to react with histidine-EGFP, and it was observed that the histidine-EGFP did selectively bind to the NTA/Ni 2+ nanoarray patterns .
  • FIG. 8 shows that a histidine-labeled fluorescence protein was applied without purification and bound to NTA/Ni nanoarray patterns.
  • histidine-EGFP protein was expressed in E. coli strain BL21 (DE3) (Novagen) .
  • the bacterial strain was separated using a centrifuge, and the supernatant containing the expressed protein was collected.
  • the cells were disrupted with, for example, an ultrasonicator and then centrifuged to collect the supernatant containing the target protein.
  • FIG. 8A shows the results of Western blotting conducted to analyze the expression of the target protein.
  • the supernatant containing the expressed target protein was allowed to react with the prepared NTA/Ni 2+ nanopatterns, and it was observed that the target protein did selectively bind to the NTA/Ni 2+ nanopatterns (see FIG. 8C) .
  • FIG. 9 shows that the target protein nanoarray can be applied not only to fluorescent proteins such as GFP (green fluorescent protein) or RFP (red fluorescent protein) , but also to various biomolecules .
  • Histidine-conjugated HRP horse radish peroxidase
  • HRP horse radish peroxidase
  • Example 2 Fabrication of His-tagged protein nanoarrays using microarrayer
  • the prior pin array or inkjet printing method has an advantage in that it is carried out using an automated process, there is a problem in that, because arrays have a size ranging from several tens to several hundreds of micrometers, either small pins capable of spotting a sample at the nanometer scale, or nozzles capable of dispensing picoliter to femtoliter volumes each time, should be fabricated in order to print spots at the nanometer scale.
  • a protein nanoarray chip was fabricated by spotting proteins using the prior microarrayer on NTA/Ni 2+ nanoarrays fabricated by the DPN technology.
  • NTA/Ni 2+ nanoarrays were fabricated, and then various histidine-protein conjugates to be immobilized were spotted on the fabricated NTA/Ni 2+ nanoarrays using a microarrayer. Washing and spotting were repeated depending on the kinds of proteins to be immobilized, thus fabricating various protein arrays on the NTA/Ni 2+ nanoarrays. It was observed that a Ni-histidine bond was selectively formed only in the NTA/Ni 2+ region among the spotted protein region. Then, the resulting nanoarrays were washed with PBS, thus fabricating a protein nanoarray chip.
  • Example 3 Fabrication of nanoarrays using mRNA-target protein fusion molecule
  • FIG. 11 shows that a fluorescence protein as a target protein was applied without purification using an mRNA- target protein fusion protein and was bound to a surface having oligonucleotide nanoarray patterns thereon.
  • a stop codon was removed from a gene coding for target protein EGFP, and the mRNA-target protein fusion molecule was expressed in vitro using a ribosome display technique.
  • the reaction solution containing the expressed target protein was allowed to react with the prepared oligonucleotide nanopatterns, and it was seen that the target protein did selectively bind to the oligonucleotide nanopatterns (see FIG. 12b) .
  • FIG. 12 shows the results of SDS-PAGE analysis of the reaction solution containing the target protein expressed in vitro. As can be seen in FIG. 12, a plurality of other proteins, in addition to the target proteins, was expressed.

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Abstract

L'invention concerne une puce à nanoréseau de protéines. Selon cette invention, les motifs des nanoréseaux de protéines sont obtenus au moyen d'un microréseau d'aiguilles ou d'un procédé de jet d'encre. Cette puce à nanoréseau de protéines peut fournir un important moyen permettant d'étudier des interactions entre des protéines associées à une maladie, dont la fonction est maintenue au simple niveau moléculaire.
PCT/KR2007/003960 2006-08-22 2007-08-20 Puce à protéines en nanoréseau et procédé de préparation de celle-ci WO2008023907A1 (fr)

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CN110389216A (zh) * 2018-04-20 2019-10-29 中国科学院化学研究所 蛋白质功能化磁珠亲和探针及其制备方法与应用

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