JP2010256032A - Resin composition for biochip production and biochip production method - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a resin composition for biochip production capable of forming a resin composition layer excellent in diffusion controllability against an acid generated by light exposure, and a method for producing a biochip using the same. <P>SOLUTION: As a result of various investigations to achieve control of diffusion of an acid generated by light exposure, it is unexpectedly found that the acid generated by light exposure is not diffused in a resin layer and selectively acts on a target first molecule by blending a radiation-sensitive acid generator and a polymer containing a nitrogen-containing group, using a nitrogen-containing organic solvent as a solvent, and a resin composition for biochip production containing (A) a polymer containing a nitrogen-containing group; (B) a radiation-sensitive acid generator; and (C) a nitrogen-containing organic solvent is completed. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

  The present invention relates to a resin composition for producing a biochip and a method for producing a biochip using the same. More specifically, the present invention relates to a biochip manufacturing resin composition used for synthesizing various polymers such as DNA, RNA, PNA and LAN on a substrate, and a biochip manufacturing method using the same.

In recent years, a method for synthesizing a polymer such as a biopolymer on a substrate has attracted attention, and in particular, a chip in which probes having different sequences and lengths are arrayed on a single substrate using nucleotides and the like as monomers, and the chip The method of manufacturing is widely studied.
As a method of synthesizing a polymer on a substrate, a nucleotide monomer having a protecting group unstable to light is arranged, and after this protecting group is dissociated from a specific portion by exposure through a mask, other nucleotides are synthesized. Patent Documents 1 and 2 disclose a method for repeating the operation of bonding monomers.
On the other hand, in the field of semiconductor manufacturing, Patent Documents 3 to 5 disclose techniques for using a photoacid generator used for forming a fine pattern using a photolithographic method and a resist containing the photoacid generator for polymer synthesis. ing.

US Pat. No. 5,445,934 US Pat. No. 5,744,305 US Pat. No. 5,658,734 JP-A-2005-0909005 Special table 2003-501640 gazette

According to the methods of Patent Documents 1 to 5, various polymers can be synthesized on a substrate. However, detection of genes and proteins using a biochip can be performed more accurately with a larger amount of specimen at a higher speed. Therefore, there is a need for a technique capable of forming more types of polymers on a substrate with higher density and accuracy than biochips.
Accordingly, an object of the present invention is to provide a resin composition for biochip production that can form a resin composition layer excellent in diffusion controllability of acid generated by exposure, and can form various types of polymers on a substrate with high density and accuracy. It is to provide a manufacturing method of a product and a biochip using the same.

  Therefore, the present inventor has made various studies to suppress the diffusion of the acid generated by exposure. When the radiation-sensitive acid generator and the polymer having a nitrogen-containing group are blended and a nitrogen-containing organic solvent is used as the solvent, Surprisingly, the present inventors have found that the acid generated by exposure does not diffuse into the resin layer and can selectively act on the target first molecule, thereby completing the present invention.

That is, the present invention comprises (A) a polymer having a nitrogen-containing group,
(B) a radiation sensitive acid generator;
(C) The resin composition for biochip manufacture containing a nitrogen-containing organic solvent is provided.

The present invention also includes (a) bonding a layer of a first molecule having an acid-labile protecting group onto a solid substrate,
(B) coating a layer of the resin composition on the layer of the first molecule;
(C) exposing the layer of the resin composition to a heat treatment to remove an acid labile protecting group from the first molecule corresponding to the exposed portion;
(D) cleaning and removing the resin composition layer from the exposed portion and the unexposed portion; and (f) binding a second molecule to the exposed first molecule. It is to provide.

  Furthermore, this invention provides the biochip formed by said manufacturing method.

According to the resin composition for producing a biochip of the present invention, a resin composition layer capable of exhibiting excellent controllability and selectivity by suppressing unnecessary diffusion of an acid generated by exposure can be formed. For this reason, by using this resin composition, it is possible to obtain a biochip with which the probe can be formed more accurately and precisely and the probe integration rate is improved as compared with the conventional case.
According to the method for producing a biochip of the present invention, a probe is formed more accurately and precisely in order to use the resin composition capable of exhibiting excellent controllability by suppressing unnecessary diffusion of acid generated by exposure. A biochip can be obtained. In addition, a biochip can be obtained in which the probe integration rate is further improved as compared with the prior art.
When at least the surface of the substrate is made of silicon, silicon dioxide, glass, polypropylene, or acrylamide, a biochip having a probe formed more accurately and precisely can be obtained.

It is explanatory drawing which illustrates the manufacturing method of the biochip of this invention typically. It is explanatory drawing which illustrates the manufacturing method of the biochip of this invention typically. It is explanatory drawing which illustrates the manufacturing method of the biochip of this invention typically.

  Hereinafter, the present invention will be described in detail. In the present specification, “(meth) acryl” means acryl and methacryl, and “(meth) acrylate” means acrylate and methacrylate.

[1] Resin composition for biochip manufacture The resin composition for biochip manufacture of the present invention contains the component (A), the component (B), and the component (C).

(1) (A) a polymer having a nitrogen-containing group (hereinafter also simply referred to as polymer (A))
The “polymer (A)” has a nitrogen-containing group. A layer obtained from an acid transfer resin composition (hereinafter also referred to as an “acid transfer resin layer”) by containing a radiation-sensitive acid generator (C) described later and having a nitrogen-containing group in the polymer (A). This layer can also be said to be an acid generator-containing layer), and can prevent unnecessary diffusion of the acid generated in the layer. For this reason, unintentional acid diffusion and acid transfer into and below the acid transfer resin layer can be prevented, and the resolution of the resulting pattern can be improved. This pattern is, for example, a pattern of a formation region of each probe in a biochip. Therefore, since these resolutions can be improved, the probes can be formed accurately and precisely, and further, the integration rate of the probes formed on the substrate can be improved. Further enhancement of functionality can be achieved in a limited area.

  As described above, the polymer (A) is preferably a polymer having substantially no acid dissociable group from the viewpoint that it preferably has an acid diffusion preventing effect and an acid transfer preventing effect. Moreover, it is preferable that a polymer (A) is a polymer which does not have a crosslinking group which bridge | crosslinks substantially by the effect | action of an acid.

  The polymer (A) is not particularly limited as long as it has a nitrogen-containing group and can suppress diffusion of the generated acid, but has an acid group, an imide group, a urea group, a urethane group, a pyridine group, and the like. The polymer which is mentioned. These nitrogen-containing groups may be present in either the main chain or the side chain, but a polymer having a nitrogen-containing group in the side chain is preferred.

  The polymer having a nitrogen-containing group in the side chain is preferably a polymer having a structural unit represented by the following general formula (1).

(In the formula, A represents a group derived from a carbon-carbon unsaturated bond; B represents a single bond or a divalent organic group; R ² and R ³ each independently represents a hydrogen atom or a substituent; Represents an optionally substituted alkyl group, an optionally substituted aryl group, an optionally substituted alicyclic hydrocarbon group or tert-butoxycarbonyl group, or R ² and R ³ may be bonded to each other to form a 3- to 10-membered monocyclic hetero ring which may further have a nitrogen atom, a sulfur atom or a selenium atom in addition to the adjacent nitrogen atom.

  Examples of the group derived from the carbon-carbon unsaturated bond represented by A in Formula (1) include a group derived from a vinyl group, a (meth) acryl group, and the like.

  Examples of the divalent organic group represented by B in the formula (1) include a divalent hydrocarbon group, and a linear or branched alkylene group having 1 to 10 carbon atoms is particularly preferable. B is preferably a single bond.

As the alkyl group which may have a substituent represented by R ¹ and R ² , a linear group having 1 to 12 carbon atoms which may be substituted with 1 to 3 of cyano group, halogen atom or the like, or A branched alkyl group can be mentioned. Examples of the alkyl group include a methyl group, an ethyl group, an n-propyl group, an isopropyl group, an n-butyl group, an isobutyl group, a tert-butyl group, an n-pentyl group, and an n-hexyl group.

  As the aryl group which may have a substituent, an aromatic hydrocarbon group having 6 to 20 carbon atoms, for example, a phenyl group, a tolyl group, a xylyl group, a medityl group, a cumenyl group, a benzyl group, a phenethyl group, a naphthyl group. Etc. These aryl groups may be substituted with 1 to 3 halogen atoms, cyano groups and the like.

  Examples of the alicyclic hydrocarbon group which may have a substituent include a monocyclic or bridged alicyclic hydrocarbon group having 3 to 12 carbon atoms, such as a cyclopentyl group, a cyclohexyl group, a bornyl group, and an isobornyl group. It is done. These alicyclic groups may be substituted with 1 to 3 alkyl groups having 1 to 4 carbon atoms, cyano groups, halogen atoms and the like.

Examples of the monocyclic heterocycle formed by combining R ¹ and R ² include pyrrolidine, hyperidine, piperazine, morpholine, thiomorpholine, and the like.

  The main chain of the polymer (A) may be a repeating unit based on a carbon-carbon unsaturated bond, and examples thereof include a group derived from a (meth) acryl group and a vinyl group, of which a (meth) acryl group The group derived from is particularly preferred.

  Preferable examples of the polymer (A) include polymers having (meth) acrylamide or (meth) acrylate having a primary amino group, secondary amino group, tertiary amino group or quaternary ammonium group in the side chain. It is done.

  A more preferable example of the polymer (A) includes a polymer having a structural unit represented by the following formula (2).

(In the formula, R ³ represents a hydrogen atom or a methyl group. R ⁴ and R ⁵ each independently represents a hydrogen atom, a linear or branched hydrocarbon group having 1 to 10 carbon atoms, or a carbon number of 3 10 to 10 cyclic hydrocarbon groups, or R ⁴ and R ⁵ may be bonded to each other and further have a nitrogen atom, an oxygen atom, a sulfur atom or a selenium atom in addition to the adjacent nitrogen atom. A 10-membered monocyclic heterocycle may be formed.)
This structural unit (2) is a structural unit derived from the monomer (Am1) represented by the following formula (3).

(In formula (3), R ³ , R ⁴ and R ⁵ are the same as above)

Examples of the linear or branched hydrocarbon group having 1 to 10 carbon atoms represented by R ⁴ and R ⁵ in the formulas (2) and (3) include a methyl group, an ethyl group, and an n-propyl group. , Alkyl groups such as i-propyl group, n-butyl group, 2-methylpropyl group, 1-methylpropyl group and t-butyl group.
That is, as the monomer (Am1) in which R ⁴ and R ⁵ in the formula (3) are linear or branched hydrocarbon groups having 1 to 10 carbon atoms, N, N-dimethyl (meth) Examples include acrylamide, N, N-diethyl (meth) acrylamide, and N-isopropyl (meth) acrylamide.

Moreover, as a C3-C10 cyclic hydrocarbon group shown by R < ⁴ > and R < ⁵ > in said Formula (2) and (3), a cyclobutyl group, a cyclopentyl group, a cyclohexyl group, a cycloheptyl group, a cyclooctyl group And alicyclic hydrocarbon groups such as phenyl group, o-tolyl group, m-tolyl group, p-tolyl group, 4-t-butylphenyl group, 1-naphthyl group, benzyl group and the like. .

Furthermore, examples of the heterocyclic ring formed by combining R ⁴ and R ⁵ in the formulas (2) and (3) include pyrrolidine, piperidine, piperazine, morpholine, thiomorpholine and the like.

  As the monomer (Am1), N, N-dimethylacrylamide, N, N-dimethylmethacrylamide, acryloylmorpholine, and methacryloylmorpholine are preferable. The resin composition of the present invention containing the polymer (A) obtained using these preferred monomers exhibits particularly excellent acid diffusion controllability. Of these, N, N-dimethylacrylamide and / or acryloylmorpholine are particularly preferable. That is, when the structural unit represented by the formula (2) contained in the polymer (A) is a structural unit derived from N, N-dialkylacrylamide or a structural unit derived from acryloylmorpholine, Diffusion can be controlled better, and it exhibits particularly excellent acid diffusion controllability.

  The polymer (A) preferably has a structural unit represented by the following formula (4) in addition to the structural unit having a nitrogen-containing group.

[In the formula (4), R ⁶ represents a hydrogen atom or a methyl group. R ⁷ represents a monovalent organic group other than an acid dissociable group. ]

  The structural unit represented by the formula (4) may be contained in the polymer (A) in any way. For example, a monomer (Am2) represented by the following formula (5) may be used. By using it, the polymer (A) containing the structural unit represented by the formula (4) can be obtained.

Wherein (5), R ⁶ and R ⁷ are as defined above. ]

The monovalent organic group having no acid-dissociable group of the R ⁷ in the formula (4) and (5), a methyl group, an ethyl group, n- propyl group, i- propyl, n- butyl group A linear or branched alkyl group having 1 to 12 carbon atoms, such as 2-methylpropyl group, 1-methylpropyl group, t-butyl group;

  Phenyl group, o-tolyl group, m-tolyl group, p-tolyl group, 2,4-xylyl group, 2,6-xylyl group, 3,5-xylyl group, mesityl group, o-cumenyl group, m-cumenyl Groups, p-cumenyl groups, benzyl groups, phenethyl groups, 1-naphthyl groups, 2-naphthyl groups and other aromatic hydrocarbon groups (especially aromatic hydrocarbon groups having 6 to 20 carbon atoms);

  Hydroxymethyl group, 1-hydroxyethyl group, 2-hydroxyethyl group, 1-hydroxypropyl group, 2-hydroxypropyl group, 3-hydroxypropyl group, 1-hydroxybutyl group, 2-hydroxybutyl group, 3-hydroxybutyl Groups, 4-hydroxybutyl groups, 3-hydroxycyclopentyl groups, 4-hydroxycyclohexyl groups and other hydroxyalkyl groups (particularly hydroxyalkyl groups having 1 to 8 carbon atoms);

  Cyano group; cyanomethyl group, 1-cyanoethyl group, 2-cyanoethyl group, 1-cyanopropyl group, 2-cyanopropyl group, 3-cyanopropyl group, 1-cyanobutyl group, 2-cyanobutyl group, 3-cyanobutyl group, 4 A nitrogen atom-containing organic group (particularly a nitrogen atom-containing organic group having 2 to 9 carbon atoms) such as a cyanobutyl group, a 3-cyanocyclopentyl group, or a 4-cyanocyclohexyl group; a cyclopentyl group, a cyclohexyl group, etc. Examples thereof include cyclic hydrocarbon groups; bridged cyclic hydrocarbon groups such as bornyl group and isobornyl group; and the like.

As the monomer (Am2), a (meth) acrylate compound is preferable. Specifically, methyl (meth) acrylate, ethyl (meth) acrylate, n-propyl (meth) acrylate, n-butyl (meth) acrylate, 2-hydroxyethyl (meth) acrylate, 2-hydroxypropyl (meth) acrylate, polyethylene glycol mono (meth) acrylate, polypropylene glycol mono (meth) acrylate, glycerol mono (meth) acrylate, phenyl (meth) acrylate, benzyl (meth) ) Acrylate, cyclohexyl (meth) acrylate, isobornyl (meth) acrylate, tricyclodecanyl (meth) acrylate, and the like. These (meth) acrylate compounds may be used alone or in combination of two or more.
Of these (meth) acrylate compounds, methyl methacrylate is particularly preferable.

  The ratio of the nitrogen-containing group-containing structural unit in the polymer (A) and the ratio of the structural unit represented by the formula (4) in the polymer (A) are not particularly limited. When the total structural unit of the union (A) is 100 mol%, the nitrogen-containing group-containing structural unit is preferably 1 to 50 mol%, more preferably 3 to 40 mol%. It is especially preferable that it is -30 mol%. When the structural unit represented by the formula (4) is included, the structural unit is preferably 5 to 99 mol%, more preferably 10 to 97 mol%, and more preferably 15 to 95 mol%. It is particularly preferred that When the proportion of the nitrogen-containing group-containing structural unit and the structural unit represented by the formula (4) in the polymer (A) is within the above range, the effect of suppressing unnecessary acid diffusion is particularly excellent. .

  The polymer (A) can further contain other structural units in addition to the nitrogen-containing group-containing structural unit and the structural unit represented by the formula (4). Other structural units include aromatic vinyl compounds such as o-hydroxystyrene, m-hydroxystyrene, p-hydroxystyrene, p-isopropenylphenol, styrene, α-methylstyrene, p-methylstyrene, and p-methoxystyrene. Cyano group-containing vinyl compounds such as acrylonitrile and methacrylonitrile; conjugated diolefins such as 1,3-butadiene and isoprene; derived from monomers such as carboxyl group-containing vinyl compounds such as acrylic acid and methacrylic acid Examples include structural units. These structural units may be used alone or in combination of two or more.

  When the other structural unit is included, the ratio is not particularly limited. However, when the total structural unit of the polymer (A) is 100 mol%, the ratio of the nitrogen-containing group-containing structural unit is 1 to 50 mol. %, More preferably 3 to 40 mol%, particularly preferably 5 to 30 mol%, and the proportion of the structural unit represented by the formula (4) is preferably 5 to 98 mol%, and 10 to 96 mol%. More preferably, 15 to 94 mol% is particularly preferable, and the proportion of other structural units is preferably 1 to 50 mol%, more preferably 1 to 30 mol%, and particularly preferably 1 to 20 mol%.

The molecular weight of the polymer (A) is not particularly limited and can be appropriately selected. The polystyrene-reduced weight molecular weight (hereinafter referred to as “Mw”) measured by gel permeation chromatography (GPC) is usually It is 1,000 to 500,000, preferably 2,000 to 400,000, and more preferably 3,000 to 300,000.
Further, the ratio (Mw / Mn) of the polymer (A) Mw and the polystyrene-equivalent molecular weight (hereinafter referred to as “Mn”) measured by GPC is not particularly limited and can be appropriately selected. Usually, it is 1-10, preferably 1-8, more preferably 1-5.

(2) (B) Radiation-sensitive acid generator The “radiation-sensitive acid generator” (hereinafter also simply referred to as “acid generator (B)”) is a component that generates an acid upon exposure. The type of radiation used for generating an acid from the acid generator (B) is not particularly limited. Ultraviolet rays, deep ultraviolet rays (including KrF excimer laser, ArF excimer laser, F ₂ excimer laser, etc.), X-rays, electrons It is appropriately selected from a line, γ-ray, molecular beam, ion beam and the like.

  The acid generator (B) is not particularly limited as long as it has an action of generating an acid upon exposure. However, onium salt compounds (including thiophenium salt compounds), halogen-containing compounds, diazoketone compounds, sulfone compounds, sulfonic acids A compound, a diazomethane compound, a sulfonimide compound, or the like can be used. This acid generator (B) may use only 1 type and may use 2 or more types together.

  Examples of the onium salt compounds include 4,7-di-n-butoxynaphthyltetrahydrothiophenium salt compounds, 1- (4-n-butoxynaphthalen-1-yl) tetrahydrothiophenium salt compounds, 1- (6- thiophenium salt compounds such as n-butoxynaphthalen-2-yl) tetrahydrothiophenium salt compound and 1- (3,5-dimethyl-4-hydroxyphenyl) tetrahydrothiophenium salt compound; bis (4-t-butylphenyl) ) Iodonium salt compounds such as iodonium salt compounds and diphenyliodonium salt compounds; triphenylsulfonium salt compounds, 4-t-butylphenyldiphenylsulfonium salt compounds, 4-cyclohexylphenyldiphenylsulfonium salt compounds, 4-methanesulfonylphenyldiphenyls Sulfonium salt compounds such as Honiumu salt compounds; phosphonium salt compound; diazonium salt compound; pyridinium salt compounds; and the like.

Examples of the halogen-containing compound include haloalkyl group-containing hydrocarbon compounds and haloalkyl group-containing heterocyclic compounds. Specific examples include (trichloromethyl) -s-triazine derivatives.
Examples of the diazoketone compound include 1,3-diketo-2-diazo compounds, diazobenzoquinone compounds, diazonaphthoquinone compounds, and the like.
Examples of the sulfonated product include β-ketosulfone, β-sulfonylsulfone, and α-diazo compounds of these compounds.
Examples of the sulfonic acid compounds include alkyl sulfonic acid esters, haloalkyl sulfonic acid esters, aryl sulfonic acid esters, and imino sulfonates.
Examples of the diazomethane compound include bis (trifluoromethylsulfonyl) diazomethane, bis (cyclohexylsulfonyl) diazomethane, bis (phenylsulfonyl) diazomethane, bis (p-toluenesulfonyl) diazomethane, methylsulfonyl-p-toluenesulfonyldiazomethane, cyclohexylsulfonyl- Examples include 1,1-dimethylethylsulfonyldiazomethane, bis (1,1-dimethylethylsulfonyl) diazomethane, and the like.

  Examples of the sulfonimide compound include N- (trifluoromethylsulfonyloxy) succinimide, N- (trifluoromethylsulfonyloxy) phthalimide, N- (trifluoromethylsulfonyloxy) diphenylmaleimide, N- (trifluoromethylsulfonyloxy). Bicyclo [2.2.1] hept-5-ene-2,3-dicarboximide, N- (trifluoromethylsulfonyloxy) -7-oxabicyclo [2.2.1] hept-5-ene-2 , 3-dicarboximide, N- (trifluoromethylsulfonyloxy) -5,6-oxy-bicyclo [2.2.1] heptane-2,3-dicarboximide, N- (trifluoromethylsulfonyloxy) Naphthylimide, N- (4-methylphenylsulfonyloxy) s Synimide, N- (4-methylphenylsulfonyloxy) phthalimide, N- (4-methylphenylsulfonyloxy) diphenylmaleimide, N- (4-methylphenylsulfonyloxy) bicyclo [2.2.1] hept-5-ene -2,3-dicarboximide, N- (4-methylphenylsulfonyloxy) -7-oxabicyclo [2.2.1] hept-5-ene-2,3-dicarboximide, N- (4- Methylphenylsulfonyloxy) -5,6-oxy-bicyclo [2.2.1] heptane-2,3-dicarboximide, N- (4-methylphenylsulfonyloxy) naphthylimide, N- (2-trifluoro) Methylphenylsulfonyloxy) succinimide, N- (2-trifluoromethylphenylsulfonyloxy) Phthalimide, N- (2-trifluoromethylphenylsulfonyloxy) diphenylmaleimide, N- (2-trifluoromethylphenylsulfonyloxy) bicyclo [2.2.1] hept-5-ene-2,3-dicarboximide N- (2-trifluoromethylphenylsulfonyloxy) -7-oxabicyclo [2.2.1] hept-5-ene-2,3-dicarboximide, N- (2-trifluoromethylphenylsulfonyloxy) ) -5,6-oxy-bicyclo [2.2.1] heptane-2,3-dicarboximide, N- (2-trifluoromethylphenylsulfonyloxy) naphthalene-1,8-dicarboximide, N- (4-Fluorophenylsulfonyloxy) succinimide, N- (4-fluorophenylsulfonyl) Ruoxy) -7-oxabicyclo [2.1.1] hept-5-ene-2,3-dicarboximide, N- (4-fluorophenylsulfonyloxy) -5,6-oxy-bicyclo [2.2 .1] Heptane-2,3-dicarboximide, N- (4-fluorophenylsulfonyloxy) naphthalene-1,8-dicarboximide, N- (10-camphor-sulfonyloxy) naphthalene-1,8-di Carboximide etc. are mentioned.

  Although content of an acid generator (B) is not specifically limited, From a viewpoint of fully ensuring the acid transferability as an acid transfer resin film, it is normally 20-100 with respect to 100 mass parts of said polymers (A). Part by mass is contained. Furthermore, from the point that excellent controllability of acid transfer by the combination of the polymer (A) and the acid generator (B) (particularly, suppression of acid diffusion in the lateral direction in the first molecular layer) can be obtained. This content is preferably 20 to 80 parts by mass, and more preferably 25 to 50 parts by mass.

(3) Nitrogen-containing organic solvent The resin composition of the present invention can freely control the state of the entire resin composition by containing a solvent, and is particularly a liquid resin composition having an arbitrary viscosity. be able to. In the present invention, it has been found that the diffusion of an acid generated by exposure can be suppressed by adding a nitrogen-containing organic solvent as the solvent.

  Examples of the nitrogen-containing organic solvent include amine-based or amide-based organic solvents. Examples of amine-based organic solvents include methylamine, ethylamine, propylamine, butylamine, pentylamine, hexylamine and aliphatic amines such as secondary and tertiary amines; aniline, N-methylaniline, N, N-dimethylaniline. , Toluidine, pyrrole, pyridine, picoline, lutidine, quinoline, isoquinoline and other aromatic amines; cyclohexylamine, dicyclohexylamine and other alicyclic amines; piperazine, piperidine, pyrrolidine, morpholine, imidazole and imidazolidinone etc. An amine; or N-methylmorpholine-N-oxide, N, N-dimethylethanolamine-N-oxide, N, N-dimethylcyclohexylamine-N-oxide, N, N, N-triethylamine-N-oxide, N, N-di Chill benzylamine -N- oxides include tertiary amine oxides such as N- methylpiperidine -N- oxide. These can be used alone or in combination.

  Examples of amide organic solvents include primary amides such as formamide, N-methylformamide, N, N-dimethylformamide, N, N-diethylformamide, acetamide, N-methylacetamide, N, N-dimethylacetamide; 2 And cyclic amides such as -pyrrolidone, N-methyl-2-pyrrolidone and 1,3-dimethyl-2-imidazolidinone. These can be used alone or in combination.

  Of these nitrogen-containing organic solvents, amide-based organic solvents are preferable, and cyclic amides are particularly preferable from the viewpoint of the effect of suppressing acid diffusion.

The resin composition of the present invention can contain another organic solvent as a solvent. As such an organic solvent,
Ketone organic solvents such as acetone, methyl ethyl ketone, methyl i-butyl ketone, cyclopentanone, cyclohexanone, 3-methylcyclopentanone, 2,6-dimethylhexanone;
Ester solvents such as ethyl acetate, n-butyl acetate, isoamyl acetate, and gamma butyrolactone;
Glycol monoether monoesters such as propylene glycol monomethyl ether acetate, propylene glycol monoethyl ether acetate, ethylene glycol monomethyl ether acetate, ethylene glycol monoethyl ether acetate, diethylene glycol monomethyl ether acetate, diethylene glycol monoethyl ether acetate;
Propylene glycol monomethyl ether, propylene glycol monoethyl ether, ethylene glycol monomethyl ether, ethylene glycol monoethyl ether, ethylene glycol dimethyl ether, ethylene glycol diethyl ether, diethylene glycol monomethyl ether, diethylene glycol monoethyl ether, diethylene glycol dimethyl ether, diethylene glycol diethyl ether, diethylene glycol ethyl methyl Ether, dipropyl ether, diisopropyl ether, butyl methyl ether, butyl ethyl ether, butyl propyl ether, dibutyl ether, diisobutyl ether, tert-butyl methyl ether, tert-butyl ethyl ether, tert-butyl propylene Ether, di-tert-butyl ether, dipentyl ether, diisoamyl ether, cyclopentyl methyl ether, cyclohexyl methyl ether, cyclopentyl ethyl ether, cyclohexyl ethyl ether, cyclopentyl propyl ether, cyclopentyl-2-propyl ether, cyclohexyl propyl ether, cyclohexyl-2- Alkyl ethers such as propyl ether, cyclopentyl butyl ether, cyclopentyl-tert-butyl ether, cyclohexyl butyl ether, cyclohexyl-tert-butyl ether;

1-propanol, n-butanol, 2-butanol, 1-pentanol, 2-pentanol, 3-pentanol, 2-methyl-1-propanol, neopentyl alcohol, tert-amyl alcohol, isoamyl alcohol, 3-methyl Alkyl alcohols such as 2-butanol, 2-methyl-1-butanol, 2-ethyl-1-butanol, 2-methyl-1-pentanol, 4-methyl-2-pentanol;
Examples include hydrocarbons such as decane, dodecane, undecane, benzene, toluene, and xylene.

  Among these organic solvents, in addition to nitrogen-containing organic solvents, ketone organic solvents and glycol monoether monoesters are used in combination to damage DNA, RNA, PNA, LAN, etc. synthesized on the substrate or acid. This is particularly preferable from the viewpoint of the diffusion suppressing effect. Further, alkyl ethers and ester solvents may be used in combination.

  The nitrogen-containing organic solvent is preferably 10 to 100% by mass, more preferably 30 to 100% by mass, and particularly preferably 40 to 90% by mass in the total organic solvent from the viewpoint of the acid diffusion inhibiting effect.

In the resin composition of the present invention, the solvent is usually contained in an amount of 10 to 10000 parts by mass, preferably 20 to 8000 parts by mass, and 30 to 6000 parts by mass, when the polymer (A) is 100 parts by mass. Is more preferable, and 40-4000 mass parts is still more preferable.
Furthermore, the viscosity of the entire resin composition in the case of containing a solvent is not particularly limited, and may be set to an appropriate viscosity for various methods for forming the acid transfer resin layer. For example, the viscosity at a temperature of 25 ° C. is 1 to 100 mPa. It can be set as s. This viscosity is preferably 2 to 80 mPa · s, and more preferably 3 to 50 mPa · s.

  The resin composition of the present invention can contain other components in addition to the above solvent. Surfactant (D) is mentioned as another component. Examples of the surfactant (D) include nonionic surfactants, anionic surfactants, cationic surfactants, amphoteric surfactants, silicone surfactants, polyalkylene oxide surfactants, and fluorine-containing surfactants. Surfactant etc. are mentioned.

Specifically, polyoxyethylene lauryl ether, polyoxyethylene stearyl ether, polyoxyethylene oleyl ether, polyoxyethylene n-octylphenyl ether, polyoxyethylene n-nonylphenyl ether, polyethylene glycol dilaurate, polyethylene glycol distearate NBX-7, NBX-8, NBX-15 (trade name, manufactured by Neos), SH8400 FLUID (trade name, manufactured by Toray Dow Corning Silicon Co.), KP341 (trade name, manufactured by Shin-Etsu Chemical Co., Ltd.) ), Polyflow No. 75, no. 95 (trade name, manufactured by Kyoeisha Chemical Co., Ltd.), F Top EF301, EF303, EF352 (trade name, manufactured by Tochem Products Co., Ltd.), Megafax F171, F172, F173, F471, R-07, R-08 (Trade name, manufactured by Dainippon Ink & Chemicals, Inc.), Florard FC430, FC431 (trade name, manufactured by Sumitomo 3M Limited), Asahi Guard AG710, Surflon S-382, SC-101, SC-102, SC-103, SC-104, SC-105, SC-106 (trade name, manufactured by Asahi Glass Co., Ltd.) and the like can be mentioned. In addition, these may use only 1 type and may use 2 or more types together.
When the surfactant (D) is used, the amount is not particularly limited, but is usually 0.01 to 0.5 parts by weight, preferably 0.02 to 100 parts by weight of the polymer (A). 0.1 parts by mass.
In addition, the resin composition of the present invention can be appropriately mixed with an acid proliferating agent, a sensitizer, a crosslinking agent, an antihalation agent, a storage stabilizer, a colorant, a plasticizer, an antifoaming agent, and the like. .

[2] Biochip production method The biochip production method of the present invention comprises:
(A) a first molecular layer forming step in which a first molecular layer composed of a first molecule having an acid-labile protecting group is directly or indirectly bonded onto a substrate;
(B) a resin composition layer forming step of forming a resin composition layer by coating the resin composition of the present invention on the first molecular layer;
(C) a protective group removing step of exposing and heat-treating the resin composition layer to remove the protective group from the first molecule constituting the first molecular layer corresponding to the exposed portion;
(D) a resin composition layer removing step for removing the resin composition layer, and
(F) a second molecule binding step of binding a second molecule to the first molecule from which the protecting group has been removed.

  In the “(a) first molecular layer forming step”, as illustrated in FIG. 1, the first molecular layer 20 composed of the first molecule having the acid-labile protecting group P is directly formed on the substrate 10. Or it is process PR1 combined indirectly.

The “first molecule” is a molecule having an acid-labile protecting group. The first molecule may be a molecule having the protective group, and the type and size thereof are not particularly limited.
Examples of the first molecule include (1) a coupling molecule (such as a compound having a protecting group and a silyl group) for directly bonding the substrate surface and the second molecule, and (2) an end of the coupling agent. Protecting group-introducing molecule for introducing a protecting group {a molecule for binding an amino group on a substrate surface treated with a coupling agent having a silyl group and an amino group to a second molecule (a peptide bond to the amino group) And the like), (3) spacer molecules for separating the substrate from the second molecule {surface of the substrate treated with a coupling agent having a silyl group and an amino group And the like (a compound having a group capable of peptide bonding to the amino group, an alkyl chain, and a protecting group).

Among these, when the coupling molecule (1) is used as the first molecule, the first molecule can be directly bonded to the substrate surface. When the protective group-introducing molecule (2) or the spacer molecule (3) is used as the first molecule, the first molecule is indirectly bonded to the substrate surface. Another coupling agent is required between one molecule and the substrate.
Among the above, the protecting group-introduced molecule (2) includes aminoalkylcarboxylic acids such as omega-aminocaproic acid compounds having a protecting group. Such compounds include 6-Nt-butoxycarbonylaminocaproic acid, 4-Nt-butoxycarbonylaminobutanoic acid, 5-Nt-butoxycarbonylaminopentanoic acid, 7-Nt-butoxycarbonyl. Examples thereof include carboxylic acid derivatives having a t-butoxycarbonyl group as a protective group such as aminoheptanoic acid.
Moreover, as a coupling rigging agent for connecting the substrate and the first molecule (protecting group-introducing molecule) when the protecting group-introducing molecule (2) is used, it has an amino group such as aminopropyltriethoxysilane and a silicon-containing group. Examples of the coupling agent include a coupling agent having a hydroxyl group and a silicon-containing group.

  In addition, as the first molecule, a compound having a protecting group among various compounds mentioned as the second molecule described later, or a derivative having a protecting group introduced into various compounds mentioned as the second molecule described later, etc. You can also.

  The “acid-labile protecting group” is a group that is dissociated by the action of an acid. This protecting group includes t-butoxycarbonyl group, dimethoxytrityl group, tetrahydropyranyl group, tetrahydrofuranyl group, (thiotetrahydropyranylsulfanyl) methyl group, (thiotetrahydrofuranylsulfanyl) methyl group, alkoxy-substituted methyl group, alkyl Examples thereof include a sulfanyl-substituted methyl group, an acetal group, a hemiacetal group, and a group represented by the following formula (6).

[In the formula (6), R ⁸ , R ⁹ and R ¹⁰ are each independently a linear or branched alkyl group having 1 to 14 carbon atoms or a non-bridged or bridged formula having 3 to 20 carbon atoms. Or a non-bridged bridge having 3 to 20 carbon atoms together with the carbon atoms to which two of R ^{8 to} R ¹⁰ are bonded to each other. A divalent alicyclic hydrocarbon group of the formula or a bridge type, and the rest is a linear or branched alkyl group having 1 to 14 carbon atoms or a non-bridged or bridged bridge having 3 to 20 carbon atoms Represents a monovalent alicyclic hydrocarbon group of the formula, each of which may be substituted. ]

  Examples of the alkoxy-substituted methyl group include methoxymethyl group, ethoxymethyl group, methoxyethoxymethyl group, n-propoxymethyl group, n-butoxymethyl group, n-pentyloxymethyl group, n-hexyloxymethyl group, and benzyl. An oxymethyl group etc. can be mentioned.

  Examples of the alkylsulfanyl-substituted methyl group include a methylsulfanylmethyl group, an ethylsulfanylmethyl group, a methoxyethylsulfanylmethyl group, an n-propylsulfanylmethyl group, an n-butylsulfanylmethyl group, an n-pentylsulfanylmethyl group, Examples thereof include an n-hexylsulfanylmethyl group and a benzylsulfanylmethyl group.

In the formula (6), examples of the linear or branched alkyl group having 1 to 14 carbon atoms of R ^{8 to} R ¹⁰ include a methyl group, an ethyl group, an n-propyl group, an I-propyl group, an n- Butyl group, 2-methylpropyl group, n-pentyl group, n-hexyl group, n-heptyl group, n-octyl group, n-nonyl group, n-decyl group, n-undecyl group, n-dodecyl group, n -A tridecyl group, n-tetradecyl group, etc. can be mentioned.

  Examples of the substituent of the alkyl group include a hydroxyl group, a carboxyl group, an oxo group (═O), a cyano group, a halogen atom (for example, a fluorine atom, a chlorine atom, etc.), a straight chain having 1 to 8 carbon atoms, Branched alkoxyl group (for example, methoxy group, ethoxy group, n-propoxy group, i-propoxy group, n-butoxy group, 2-methylpropoxy group, 1-methylpropoxy group, t-butoxy group, etc.), carbon number 2-8 linear or branched alkoxyalkoxyl groups (for example, methoxymethoxy group, ethoxymethoxy group, t-butoxymethoxy group, etc.), C2-C8 linear or branched alkylcarbonyloxy group (For example, methylcarbonyloxy group, ethylcarbonyloxy group, t-butylcarbonyloxy group, etc.), straight chain having 2 to 8 carbon atoms Or branched alkoxycarbonyl group (e.g., methoxycarbonyl group, ethoxycarbonyl group, t-butoxycarbonyl group) can be exemplified one or more, or one or more such.

Examples of the non-bridged or bridged monovalent alicyclic hydrocarbon group of R ^{8 to} R ¹⁰ in formula (6) include a cyclopropyl group, a cyclobutyl group, and a cyclopentyl group. Group, a cycloalkyl group such as a cyclohexyl group, a cycloheptyl group, and a cyclooctyl group; a bicyclo [2.2.1] heptyl group, a bicyclo [2.2.2] octyl group, a tetracyclo [4.2.0.1 ^{2, 5} . 1 ^7,10 ] dodecyl group, adamantyl group and the like.

Examples of the monovalent alicyclic hydrocarbon groups and substituents of the divalent alicyclic hydrocarbon group two of them formed by binding to each other of R ⁸ to R ¹⁰ of formula (6) , For example, a hydroxyl group, a carboxyl group, an oxo group (═O), a cyano group, a halogen atom (for example, a fluorine atom, a chlorine atom, etc.), a linear or branched alkyl group having 1 to 14 carbon atoms (for example, Methyl group, ethyl group, n-propyl group, i-propyl group, n-butyl group, 2-methylpropyl group, 1-methylpropyl group, t-butyl group, etc.), straight chain having 1 to 8 carbon atoms or Branched alkoxyl group (for example, methoxy group, ethoxy group, n-propoxy group, i-propoxy group, n-butoxy group, 2-methylpropoxy group, 1-methylpropoxy group, t-butoxy group, etc.), carbon number 2 to 8 linear young Or a branched alkoxyalkyl group (for example, methoxymethyl group, ethoxymethyl group, t-butoxymethyl group, etc.), a linear or branched alkoxyalkoxyl group having 2 to 8 carbon atoms (for example, methoxymethoxy group, Ethoxymethoxy group, t-butoxymethoxy group, etc.), C2-C8 linear or branched alkylcarbonyloxy group (for example, methylcarbonyloxy group, ethylcarbonyloxy group, t-butylcarbonyloxy group, etc.) , A linear or branched alkoxycarbonyl group having 2 to 8 carbon atoms (for example, methoxycarbonyl group, ethoxycarbonyl group, t-butoxycarbonyl group, etc.), linear or branched cyano having 2 to 14 carbon atoms An alkyl group (for example, a cyanomethyl group, a 2-cyanoethyl group, a 3-cyanopropyl group, -Cyanobutyl group, etc.), 1 or more kinds or 1 or more kinds of C1-C14 linear or branched fluoroalkyl groups (for example, fluoromethyl group, trifluoromethyl group, pentafluoroethyl group, etc.) Can be mentioned.

  The kind of the “substrate” is not particularly limited, and may be made of an inorganic material, an organic material, or a composite material thereof. The substrate may be made of different materials on the front side and the other side. Examples of the substrate material include inorganic materials whose main component is silicon, such as silicon, silicon dioxide, and glass (including borosilicate glass, surface-modified glass, and quartz glass). Further, organic materials such as polypropylene and acrylamide (including activated acrylamide) can be mentioned. In addition, other substrates known in the art having a surface having a reactive site suitable for immobilizing a layer of a molecule having an unstable protecting group can be appropriately used.

  The first molecular layer may be bonded on the substrate in any way, but usually, a liquid containing the first molecule is applied to the surface of a substrate (including a substrate not subjected to surface treatment and a substrate subjected to surface treatment). Then, the first molecule and the substrate surface are reacted and bonded. The coating method in this case is not particularly limited, and various conventionally known methods such as spin coating, cast coating, roll coating, and printing can be used.

  In the “(b) resin composition layer forming step”, as illustrated in FIG. 1, the first molecular layer 20 is coated with the resin composition of the present invention (that is, the acid transfer resin composition). This is a process PR2 for forming the resin composition layer (that is, the acid transfer resin layer) 30.

The coating means for the acid transfer resin composition is not particularly limited, and examples thereof include appropriate application means such as spin coating, cast coating, roll coating, and printing.
Furthermore, after apply | coating this acid transfer resin composition, you may form an acid transfer resin layer by volatilizing the solvent in a coating film by pre-baking (PB) as needed. The heating conditions for this prebaking are appropriately selected depending on the composition of the acid transfer resin composition, but the heating temperature is usually about 30 to 150 ° C, preferably 50 to 130 ° C. Furthermore, the heating time is usually 30 to 300 seconds, preferably 60 to 180 seconds.
The thickness of the acid transfer resin layer is not particularly limited, but is usually preferably 1 to 10000 nm, more preferably 5 to 800 nm, and still more preferably 10 to 500 nm.

  In the “(c) protecting group removing step”, as illustrated in FIGS. 1 and 2, the acid transfer resin layer 30 is exposed and heat-treated to form the first molecular layer 30 corresponding to the exposed portion. Steps PR3 and PR4 for removing the protecting group P from the first molecule. In this protective group removing step, usually, an exposure step PR3 for exposing the acid transfer resin layer 30 to radiation, and an acid generated in the acid transfer resin layer 30 by exposure to the first molecular layer 20 are transferred (diffused). And a transfer step PR4.

Of these, the exposure step PR3 is a step of exposing the acid transfer resin layer 30 through the mask 50 to generate an acid in the acid transfer resin layer 30. As a result, as illustrated in FIG. 1, the exposed portion of the acid transfer resin layer 30 becomes the acid generation portion 31.
The type of radiation used for the exposure is not particularly limited, and ultraviolet rays, far ultraviolet rays (KrF excimer laser, ArF excimer laser, F ₂ excimer) are selected depending on the type of acid generator (B) contained in the acid transfer resin layer 30. (Including laser etc.), X-ray, electron beam, γ-ray, molecular beam, ion beam and the like. Further, the exposure amount and the like are appropriately selected according to the type of the acid generator (B) contained in the acid transfer resin layer 30.

The acid transfer process PR4 is a process of transferring the acid generated in the acid transfer resin layer 30 to the first molecular layer 20. As a result, as illustrated in FIG. 2, a part of the first molecular layer 20 corresponding to the acid generation site 31 becomes an acid transfer site 21 (a site composed of the residue of the first molecule from which the protecting group is dissociated).
The method for transferring the acid is not particularly limited. Specifically, (1) a method for transferring by heating, (2) a method for transferring by standing at room temperature, and (3) a transfer using osmotic pressure. The method etc. are mentioned. These methods may be used alone or in combination of two or more. Among these methods, (1) the method of transferring by heating is preferable because of excellent transfer efficiency.
The heating conditions for transferring by heating are not particularly limited, but the heating temperature is preferably 50 to 200 ° C, more preferably 70 to 150 ° C. Furthermore, the heating time is preferably 30 to 300 seconds, more preferably 60 to 180 seconds.
In addition, when transferring by heating, it may be completed by one heating depending on the above heating conditions, but as a result, two or more heatings may be performed so as to obtain the same result as the above heating conditions. it can.

In addition, the method of transferring by leaving at (2) normal temperature is not generated by heating, and is usually generated in the acid transfer resin layer 30 by leaving in a normal temperature environment of 20 to 30 ° C. In this method, the acid is naturally diffused and transferred to the first molecular layer 20.
Furthermore, (3) the method of transferring using osmotic pressure is the difference between the osmotic pressure of the acid component between the acid transfer resin layer 30 and the first molecular layer 20 by using the acid concentration difference. This is a transfer method in which the acid in the acid transfer resin layer 30 is diffused to the first molecular layer 20 at a diffusion rate higher than that of natural diffusion.

The “(d) resin composition layer removing step” is a step PR5 of removing the resin composition layer 30 as illustrated in FIG. That is, it is a step of removing the acid transfer resin layer 30 and exposing the first molecular layer 20 to which the acid has been transferred under the layer.
The acid transfer resin layer 30 may be removed by any method, but the acid transfer resin layer 30 is usually dissolved in an organic solvent. This organic solvent dissolves the acid transfer resin layer 30 but does not dissolve the first molecular layer 20 to which the acid has been transferred.

  Such an organic solvent is preferably selected as appropriate depending on the resin composition of the acid transfer resin layer 30 and the first molecular layer 20, and is an organic in which the first molecular layer 20 is not dissolved and the acid transfer resin layer 30 is dissolved. Although it will not be limited if it is a solvent, Specifically, acetonitrile, acetone, tetrahydrofuran, a pyridine, etc. are mentioned. These organic solvents may use only 1 type and may use 2 or more types together.

  As illustrated in FIG. 2, the “(f) second molecule bonding step” includes the first molecule from which the protecting group P has been removed (the first molecule that has been bonded to the substrate 10 and from which the protecting group P has been removed). This is a step PR6 in which the second molecule is bound to the residue of the molecule. That is, in the first molecular layer 20, a portion 41 made of the second molecule is stacked on the portion 21 where the acid transfer is performed and the protecting group P of the first molecule is dissociated.

  The type of the “second molecule” is not particularly limited, and various molecules can be used. Examples of the second molecule include (1) nucleotides {including nucleotides, deoxynucleotides and analogs excluding these (synthetic nucleotide analogs, synthetic deoxynucleotide analogs, etc.)}, (2) amino acids, (3) single Saccharides, or (4) a conjugate in which two or more molecules selected from these nucleotides, amino acids and monosaccharides are combined, and (5) a peptide nucleic acid-forming molecule for synthesizing peptide nucleic acid (PNA) (peptide nucleic acid monomer). ) And (6) various end-forming molecules. These second molecules may have a protecting group and an active group. Moreover, these 2nd molecules may use only 1 type, and may use 2 or more types together.

Examples of the nucleotide (1) include nucleotides, deoxynucleotides, and synthetic nucleotide analogs.
Among these, examples of the nucleotide include adenosine phosphate, guanosine phosphate, cytidine phosphate, uridine phosphate and the like.
Examples of deoxynucleotides include deoxyadenosine phosphate, deoxyguanosine phosphate, deoxythidine phosphate, deoxythymidine phosphate, and the like.
Furthermore, the synthetic nucleotide analogs include 2′-4 ′ cross-linked nucleotide analogs, 3′-4 ′ cross-linked nucleotide analogs, 5′-amino-3 ′, 5 ′ cross-linked nucleotide analogs and the like. Etc.

Examples of (2) amino acids (including L-form and D-form) include glycine, alanine, valine, leucine, isoleucine, serine, threonine, cysteine, methionine, asparagine, glutamine, proline phenylalanine, tyrosine, tryptophan and the like.
Examples of the (3) monosaccharide include glucose, galactose, mannose, fucose, xylose, N-acetylglucosamine, N-acetylgalactosamine and the like.
Examples of the conjugate of (4) include oligonucleotides that are conjugates of nucleotides, peptides and proteins that are conjugates of amino acids.

  Examples of the peptide nucleic acid-forming molecule include N- (2-t-butyloxycarbonyl-aminoethyl) -N-thymin-1-ylacetyl) glycine, N- (N-4- (benzyloxycarbonyl) cytosine-1- Yl) acetyl-N- (2-t-butyloxycarbonyl-aminoethyl) glycine, N- (N-6- (benzyloxycarbonyl) adenine-9-yl) acetyl-N- (2-t-butyloxycarbonyl) -Aminoethyl) glycine and N- (N-4- (benzyloxycarbonyl) guanin-1-yl) acetyl-N- (2-t-butyloxycarbonyl-aminoethyl) glycine.

  The (5) end-forming molecule is a molecule that forms a molecular chain end, and includes a protecting group-forming molecule having various protecting groups, various capping molecules, a labeling molecule, and the like. Among these, the labeling molecules include various fluorescent labeling compounds (fluorescein derivatives such as phloresin isothiocyanate) and radioisotope labeling compounds.

  Furthermore, as the protecting group that the second molecule can have, the acid-labile protecting group in the first molecule can be applied as it is, and a light-labile protecting group can also be used.

  Examples of the active group that the second molecule can have include phosphorus-containing groups that can react with free hydroxyl groups such as phosphoramidite groups, H-phosphonates, phosphodiesters, phosphotriesters, and phosphate triesters. It is done. Thus, for example, activated nucleotides include phosphoramidite nucleotide molecules. In addition, examples of the photochemically active group and the thermochemically active group include an amino group, a thiol group, a maleimide group, an N-hydroxysuccinimidyl ester group, a formyl group, a carboxyl group, an acrylamide group, and an epoxy group.

  Then, as illustrated in FIG. 3, the same operations as those for dissociating the protecting group from the first molecule described above (resin composition layer formation step PR7, exposure step PR8, acid transfer step PR9, resin composition layer removal) By applying the step PR10), the protecting group is dissociated from the remaining first molecule (the second molecule is not bound), and then the third molecule binding step PR11 is performed to thereby obtain the residue of the first molecule. A third molecule can be bound to to form a site 42 consisting of residues of the third molecule.

Furthermore, as illustrated in the bottom diagram of FIG. 2, when the second molecule has an acid-labile protecting group P, the second molecule consists of residues of the second molecule by performing the same operation as described above. Other molecules (fourth molecule, fifth molecule, etc.) can be bound on the site 41. By repeating the same operation in this way, a polymer can be synthesized with a high degree of freedom on the substrate.
The description relating to the second molecule can be applied to the third molecule, the fourth molecule and the fifth molecule as they are. In addition, the first molecule, the second molecule, the third molecule, the fourth molecule, and the fifth molecule may be the same or different.

  According to the production method of the present invention, a polymer can be designed with a high degree of freedom on a biochip substrate. The polymer synthesized by this method is not particularly limited, but is particularly suitable for synthesis of biopolymers and pseudo-biopolymers. Examples of such a polymer include nucleic acids and proteins. Examples of nucleic acids include DNA, RNA and PNA (Peptide Nucleic Acid), as well as artificial nucleic acids (LNA {Locked Nucleic Acid (trademark of Proligo LLC)}) and BNA synthesized using partially or fully cross-linked nucleotide analogs. Etc.]. Among these, PNA is a pseudo-biopolymer having a peptide bond skeleton, whereas DNA and RNA have a phosphate bond skeleton. This PNA is usually a polymer having an aminoethylglycine derivative as a monomer.

  Hereinafter, the embodiment of the present invention will be described more specifically with reference to examples. However, the present invention is not limited to this embodiment. In the description of Examples, “part” and “%” are based on mass unless otherwise specified.

[1] Preparation of resin composition (A1 and A2)
(1) Synthesis of Polymer (A) [Synthesis Example 1] <Synthesis of Polymer A1>
In this synthesis example 1, N, N-dimethylacrylamide represented by the following formula (7) is used as the monomer (Am1) for introducing the structural unit represented by the formula (2). In this example, methyl methacrylate is used as a monomer (Am2) for introducing the structural unit represented by 4).

In a 500 mL beaker, 5 g of N, N-dimethylacrylamide (monomer Am1, manufactured by Kojin Co., Ltd.) (5 mol% when the total of Am1 and Am2 is 100 mol%), methyl methacrylate (monomer Am2) , Manufactured by Mitsubishi Materials Corporation), 95 g (95 mol% when the total of Am1 and Am2 is 100 mol%), and 3.0 g of 2,2′-azobisisobutyronitrile (polymerization initiator) The mixture was stirred until the polymerization initiator was dissolved to obtain a uniform solution. Separately, 150 g of propylene glycol monomethyl ether acetate (solvent) was charged into a flask equipped with a nitrogen-substituted dry ice / methanol refluxer, gently stirred and heated to 80 ° C. Then, the said solution was dripped little by little over 2 hours at 80 degreeC. After the dropping, polymerization was further performed at 80 ° C. for 3 hours, and then the temperature was raised to 100 ° C. and stirred for 1 hour to complete the polymerization. Thereafter, the obtained reaction solution was dropped into a large amount of cyclohexane to solidify the product. Next, the obtained coagulated product was washed with water, redissolved in tetrahydrofuran having the same mass as the coagulated product, and dropped into a large amount of cyclohexane to be coagulated again. After performing this re-dissolution and solidification cycle three times in total, the obtained solidified product was vacuum dried at 40 ° C. for 48 hours to obtain a polymer A1.
The yield of the obtained polymer A1 was 90%, Mw was 11,000, and Mw / Mn was 2.3. The polymer A1 is a molar (5:95) polymer having both the structural unit represented by the formula (2) and the structural unit represented by the formula (4).

[Synthesis Example 2] <Synthesis of Polymer A2>
In Synthesis Example 2, 10 g of N, N-dimethylacrylamide (monomer Am1, manufactured by Kojin Co., Ltd.) in Synthesis Example 1 (10 mol% when the total of Am1 and Am2 is 100 mol%) , 90 g (monomer Am2, manufactured by Mitsubishi Materials Co., Ltd.) (90 mol% when the total of Am1 and Am2 is 100 mol%) It was.
Mw of the obtained polymer A2 was 10,000. The polymer A2 is a polymer having both the structural unit represented by the formula (2) and the structural unit represented by the formula (4).

(2) Mixing of each component Polymer A (100 parts by mass) so that the polymer A (A1 and A2) and the following three components (acid generator, solvent and surfactant) are blended as shown in Table 2 below. ), Acid generator B (30 parts by mass), solvent C (1300 to 1350 parts by mass), and surfactant D (0.05 parts by mass) are mixed and stirred to form a uniform solution (each solid shown in Table 2). Minute concentration). This solution was filtered through a capsule filter having a pore size of 0.5 μm to obtain 7 types of resin compositions (Examples 1 to 5 and Comparative Examples 1 and 2).

  As the acid generator (B), N- (methanesulfonyloxy) naphthalene-1,8-dicarboximide (B1) (Midori Chemical Co., Ltd. product name “NAI-100”), or 4,7-di-n- Butoxynaphthyltetrahydrothiophenium trifluoromethanesulfonate (B2) (manufactured by ADEKA Corporation) was used.

Solvent C1-1: N-methyl-2-pyrrolidone C2-1: Cyclohexanone C3-1: Gamma butyrolactone C3-2: Propylene glycol monomethyl ether acetate

  As the surfactant (D), trade name “Dynaflow” manufactured by JSR Corporation was used.

[2] Evaluation of Resin Composition After selectively removing protecting groups of the first molecular layer using each resin composition in order to evaluate the characteristics of each resin composition obtained in [1]. Then, fluorescent labeling was performed, and the shape of each spot was evaluated.

(1) First molecular layer forming step The glass substrate was immersed in a cleaning solution (1 L of 95% ethanol aqueous solution, 12 mL of water, 120 g of sodium hydroxide) for 12 hours, then washed with water several times and dried in the air. Next, a surface treatment for fixing amino groups on the glass substrate was performed. That is, the glass substrate was immersed in an ethanol solution of 0.1% by volume aminopropyltriethoxysilane and stirred at room temperature for 5 minutes. Then, it was washed with ethanol three times, dried at 120 ° C. for 20 minutes using a vacuum oven, and further left in an argon gas atmosphere for 12 hours, and then N, N-dimethylformamide (hereinafter simply referred to as “DMF”). Then, the surface treatment was performed by washing with dichloromethane.

  Thereafter, the surface-treated glass substrate was added to 0.5 mL of a DMF solution containing 30 mM 6-Nt-butoxycarbonylaminocaproic acid (first molecule in this example) and 3 g of dicyclohexylcarbodiimide (DCC). It was immersed and reacted at 80 ° C. with stirring for 1 hour. Then, in order to protect an unreacted amino group with an acetyl group, it was made to react, stirring for 1 hour in the mixed solution (1 volume part of acetic anhydride +3 volume part of pyridine) of acetic anhydride and pyridine. As a result, a first molecular layer (linker layer) composed of a first molecule in which the amino group was protected with an acid-labile protecting group (acetyl group) was formed on the glass substrate.

(2) Resin composition layer forming step Each resin composition (Examples 1 to 5 and Comparative Examples 1 and 2) obtained in [1] is replaced with the first molecule obtained in [2] (1). The glass substrate on which the layer was formed was coated using a spin coater and then heated on a hot plate at 110 ° C. for 1 minute to form each resin composition layer having a thickness of 150 nm.

(3) Protecting group removal step An ultrahigh pressure mercury lamp (manufactured by OSRAM Co., Ltd., type) is formed on the surface of the resin composition layer of the glass substrate obtained up to (2) above through a pattern mask (square pattern of 50 μm × 50 μm). Using “HBO” (output: 1,000 W), ultraviolet light of 100 to 1000 mJ / cm ² was irradiated to generate an acid in the resin composition layer. The exposure amount was confirmed by an illuminance meter [manufactured by Oak Manufacturing Co., Ltd., type “UV-M10” (illuminance meter) connected to type “probe UV-35” (receiver)].
Next, the glass substrate after the exposure was again heated on a hot plate at 110 ° C. for 1 minute to transfer the acid generated in the resin composition layer to the first molecular layer.

(4) Resin composition layer removal process The glass substrate obtained by said (3) was immersed in acetonitrile for 30 second, and the said resin composition layer was removed.

(5) Second molecule binding step A protective group is dissociated from the first molecule in the step (3) and is exposed to the glass substrate surface in the step (4). In a DMF solution containing 1 mM phloresin isothiocyanate (manufactured by Aldrich, second molecule in the present example), a fluorescent label was formed by reacting at room temperature for 1 hour. Then, after wash | cleaning in order of ethanol, water, and ethanol, it was made to dry and stored in the dark room.

(6) Evaluation of spot shape While observing the glass substrate obtained up to (5) above using a microscopic laser Raman spectrometer (manufactured by Renishaw), the shape of each spot is evaluated based on the following criteria, The results are shown in Table 3 below.
“◯”: When the absorption of the isothiocyanate group is observed uniformly over the entire surface of the 50 μm × 50 μm square, and the absorption of the isothiocyanate group is observed outside the square.
“Δ”: When absorption of an isothiocyanate group is observed in a part of a square of 50 μm × 50 μm and absorption of an isothiocyanate group is observed outside the square.
“×”: When absorption of an isothiocyanate group is observed both inside and outside the square of 50 μm × 50 μm.

  In addition, in this invention, it can restrict to what is shown to said specific Example, It can be set as the Example variously changed within the range of this invention according to the objective and the use.

10; substrate,
20; first molecular layer, 21; site where the protecting group is dissociated, P: protecting group,
30; Resin composition layer (acid transfer resin layer), 31; Acid generation site,
41; site consisting of residues of the second molecule, 42; site consisting of residues of the third molecule (the other second molecule),
50; mask,
PR1; first molecular layer forming step, PR2; resin composition layer (acid transfer resin layer) forming step, PR3; exposure step (part of protecting group removing step), PR4; acid transferring step (one protecting group removing step) Part), PR5; resin composition layer (acid transfer resin layer) removal step, PR6; second molecule binding step,
PR7; resin composition layer (acid transfer resin layer) formation step, PR8; exposure step, PR9; acid transfer step, PR10; resin composition layer (acid transfer resin layer) removal step, PR11; third molecule binding step.

Claims (13)

(A) a polymer having a nitrogen-containing group;
(B) a radiation sensitive acid generator;
(C) A resin composition for producing biochips containing a nitrogen-containing organic solvent.   The resin composition for producing biochips according to claim 1, wherein the component (A) is a polymer having a nitrogen-containing group in the side chain.   (B) Content of a component is 20-100 mass parts with respect to 100 mass parts of (A) component, The resin composition for biochip manufacture of Claim 1 or 2.   The resin composition for biochip production according to any one of claims 1 to 3, wherein the nitrogen-containing organic solvent (C) is an amide organic solvent.   Furthermore, the resin composition for biochip manufacture of any one of Claims 1-4 containing ketone type | system | group organic solvent or glycol monoether monoesters.   Content of the said nitrogen-containing organic solvent (C) is 10-100 mass% in the organic solvent in the said composition, The resin composition for biochip manufacture of any one of Claims 1-5. (A) A component is a polymer which has a structural unit shown by General formula (1), The resin composition for biochip manufacture of any one of Claims 1-6.

(In the formula, A represents a group derived from a carbon-carbon unsaturated bond; B represents a single bond or a divalent organic group; R ¹ and R ² each independently represents a hydrogen atom or a substituent; Represents an optionally substituted alkyl group, an optionally substituted aryl group, an optionally substituted alicyclic hydrocarbon group or tert-butoxycarbonyl group, or R ¹ and R ² may be bonded to each other to form a 3- to 10-membered monocyclic heterocyclic ring which may further have a nitrogen atom, a sulfur atom or a selenium atom in addition to the adjacent nitrogen atom. (A) The component is a polymer which has a structural unit shown by following formula (2), The resin composition for biochip manufacture of any one of Claims 1-7.

(In the formula, R ³ represents a hydrogen atom or a methyl group. R ⁴ and R ⁵ each independently represents a hydrogen atom, a linear or branched hydrocarbon group having 1 to 10 carbon atoms, or a carbon number of 3 10 to 10 cyclic hydrocarbon groups, or R ⁴ and R ⁵ may be bonded to each other and further have a nitrogen atom, an oxygen atom, a sulfur atom or a selenium atom in addition to the adjacent nitrogen atom. A 10-membered monocyclic heterocycle may be formed.) The resin composition for biochip production according to claim 8, wherein the polymer having the structural unit represented by the formula (2) further has a structural unit represented by the following formula (4).

[In the formula (4), R ⁶ represents a hydrogen atom or a methyl group. R ⁷ represents a monovalent organic group other than an acid dissociable group. ] (A) attaching a layer of a first molecule having an acid labile protecting group to a solid substrate;
(B) coating the layer of the resin composition according to any one of claims 1 to 9 on the layer of the first molecule;
(C) exposing the layer of the resin composition to a heat treatment to remove an acid labile protecting group from the first molecule corresponding to the exposed portion;
(D) A method for producing a biochip, comprising: washing and removing a resin composition layer from the exposed part and the unexposed part; and (f) binding a second molecule to the exposed first molecule.   The method for producing a biochip according to claim 10, wherein at least a surface of the solid substrate is made of silicon, silicon dioxide, glass, surface-modified glass, polypropylene, or activated acrylamide.   The method for producing a biochip according to claim 10 or 11, wherein the second molecule is a monomer of a nucleic acid or a protein.   The biochip formed by the manufacturing method of the biochip of any one of Claims 10-12.

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