CN106872425A - A kind of modularization biological sample analysis chip - Google Patents
A kind of modularization biological sample analysis chip Download PDFInfo
- Publication number
- CN106872425A CN106872425A CN201710023509.1A CN201710023509A CN106872425A CN 106872425 A CN106872425 A CN 106872425A CN 201710023509 A CN201710023509 A CN 201710023509A CN 106872425 A CN106872425 A CN 106872425A
- Authority
- CN
- China
- Prior art keywords
- chip
- analysis chip
- biological sample
- molecule
- analysis
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 238000004458 analytical method Methods 0.000 title claims abstract description 139
- 239000012472 biological sample Substances 0.000 title claims abstract description 39
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 109
- 239000000463 material Substances 0.000 claims abstract description 87
- 239000000758 substrate Substances 0.000 claims abstract description 49
- 238000006243 chemical reaction Methods 0.000 claims abstract description 19
- 238000001179 sorption measurement Methods 0.000 claims abstract description 13
- 230000002209 hydrophobic effect Effects 0.000 claims abstract description 11
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 7
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 7
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 7
- 238000012351 Integrated analysis Methods 0.000 claims abstract description 6
- 229920001184 polypeptide Polymers 0.000 claims abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 6
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 6
- 238000004140 cleaning Methods 0.000 claims description 26
- 238000005520 cutting process Methods 0.000 claims description 13
- 238000001514 detection method Methods 0.000 claims description 12
- 230000027455 binding Effects 0.000 claims description 8
- 239000011521 glass Substances 0.000 claims description 8
- 230000004048 modification Effects 0.000 claims description 8
- 238000012986 modification Methods 0.000 claims description 8
- 102000004190 Enzymes Human genes 0.000 claims description 7
- 108090000790 Enzymes Proteins 0.000 claims description 7
- 239000004593 Epoxy Substances 0.000 claims description 7
- 125000000217 alkyl group Chemical group 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 7
- 238000005859 coupling reaction Methods 0.000 claims description 7
- 239000002096 quantum dot Substances 0.000 claims description 7
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 claims description 6
- -1 amino, chloromethyl Chemical group 0.000 claims description 6
- 230000008878 coupling Effects 0.000 claims description 6
- 238000010168 coupling process Methods 0.000 claims description 6
- 239000004033 plastic Substances 0.000 claims description 6
- 229920003023 plastic Polymers 0.000 claims description 6
- 239000004677 Nylon Substances 0.000 claims description 5
- 239000004793 Polystyrene Substances 0.000 claims description 5
- 235000001014 amino acid Nutrition 0.000 claims description 5
- 150000001413 amino acids Chemical class 0.000 claims description 5
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 claims description 5
- 239000007769 metal material Substances 0.000 claims description 5
- 229920001778 nylon Polymers 0.000 claims description 5
- 229920000136 polysorbate Polymers 0.000 claims description 5
- 229920002223 polystyrene Polymers 0.000 claims description 5
- 235000000346 sugar Nutrition 0.000 claims description 5
- 230000004913 activation Effects 0.000 claims description 4
- 239000005018 casein Substances 0.000 claims description 4
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims description 4
- 235000021240 caseins Nutrition 0.000 claims description 4
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 claims description 3
- 229960002685 biotin Drugs 0.000 claims description 3
- 235000020958 biotin Nutrition 0.000 claims description 3
- 239000011616 biotin Substances 0.000 claims description 3
- 229920002301 cellulose acetate Polymers 0.000 claims description 3
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 claims description 3
- 238000012284 sample analysis method Methods 0.000 claims description 3
- 239000002210 silicon-based material Substances 0.000 claims description 3
- 235000020183 skimmed milk Nutrition 0.000 claims description 3
- 239000000811 xylitol Substances 0.000 claims description 3
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 claims description 3
- 229960002675 xylitol Drugs 0.000 claims description 3
- 235000010447 xylitol Nutrition 0.000 claims description 3
- 229910052751 metal Inorganic materials 0.000 claims description 2
- 239000002184 metal Substances 0.000 claims description 2
- 102000003886 Glycoproteins Human genes 0.000 claims 1
- 108090000288 Glycoproteins Proteins 0.000 claims 1
- 239000008280 blood Substances 0.000 claims 1
- 210000004369 blood Anatomy 0.000 claims 1
- 239000000523 sample Substances 0.000 abstract description 45
- 238000000034 method Methods 0.000 abstract description 20
- 230000008569 process Effects 0.000 abstract description 11
- 230000009871 nonspecific binding Effects 0.000 abstract description 5
- 239000007788 liquid Substances 0.000 description 20
- PXIPVTKHYLBLMZ-UHFFFAOYSA-N Sodium azide Chemical compound [Na+].[N-]=[N+]=[N-] PXIPVTKHYLBLMZ-UHFFFAOYSA-N 0.000 description 12
- 239000011248 coating agent Substances 0.000 description 7
- 238000000576 coating method Methods 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 102000004169 proteins and genes Human genes 0.000 description 7
- 108090000623 proteins and genes Proteins 0.000 description 7
- 239000007853 buffer solution Substances 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 238000003698 laser cutting Methods 0.000 description 6
- 238000012360 testing method Methods 0.000 description 6
- 238000000018 DNA microarray Methods 0.000 description 5
- DZBUGLKDJFMEHC-UHFFFAOYSA-N benzoquinolinylidene Natural products C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 230000005284 excitation Effects 0.000 description 4
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 3
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 108090001008 Avidin Proteins 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 2
- 239000004472 Lysine Substances 0.000 description 2
- 229920005654 Sephadex Polymers 0.000 description 2
- 239000012507 Sephadex™ Substances 0.000 description 2
- XUIMIQQOPSSXEZ-UHFFFAOYSA-N Silicon Chemical compound [Si] XUIMIQQOPSSXEZ-UHFFFAOYSA-N 0.000 description 2
- 108010090804 Streptavidin Proteins 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000007413 biotinylation Methods 0.000 description 2
- 229940098773 bovine serum albumin Drugs 0.000 description 2
- 239000006172 buffering agent Substances 0.000 description 2
- 230000003139 buffering effect Effects 0.000 description 2
- 238000005119 centrifugation Methods 0.000 description 2
- 239000003431 cross linking reagent Substances 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 150000004985 diamines Chemical class 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 150000004676 glycans Chemical class 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 239000004816 latex Substances 0.000 description 2
- 229920000126 latex Polymers 0.000 description 2
- 238000004020 luminiscence type Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- RXNXLAHQOVLMIE-UHFFFAOYSA-N phenyl 10-methylacridin-10-ium-9-carboxylate Chemical compound C12=CC=CC=C2[N+](C)=C2C=CC=CC2=C1C(=O)OC1=CC=CC=C1 RXNXLAHQOVLMIE-UHFFFAOYSA-N 0.000 description 2
- 229920001282 polysaccharide Polymers 0.000 description 2
- 239000005017 polysaccharide Substances 0.000 description 2
- 230000037452 priming Effects 0.000 description 2
- 239000010703 silicon Substances 0.000 description 2
- 229910052710 silicon Inorganic materials 0.000 description 2
- RPENMORRBUTCPR-UHFFFAOYSA-M sodium;1-hydroxy-2,5-dioxopyrrolidine-3-sulfonate Chemical compound [Na+].ON1C(=O)CC(S([O-])(=O)=O)C1=O RPENMORRBUTCPR-UHFFFAOYSA-M 0.000 description 2
- PXQLVRUNWNTZOS-UHFFFAOYSA-N sulfanyl Chemical class [SH] PXQLVRUNWNTZOS-UHFFFAOYSA-N 0.000 description 2
- 230000000153 supplemental effect Effects 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- AYLWQNNBQZVDHD-UHFFFAOYSA-N 1,2-dimethylacridine nitric acid Chemical compound [N+](=O)(O)[O-].CC1=C(C2=CC3=CC=CC=C3N=C2C=C1)C AYLWQNNBQZVDHD-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- RCEAADKTGXTDOA-UHFFFAOYSA-N OS(O)(=O)=O.CCCCCCCCCCCC[Na] Chemical compound OS(O)(=O)=O.CCCCCCCCCCCC[Na] RCEAADKTGXTDOA-UHFFFAOYSA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- BDQRMEBGHYKVLA-UHFFFAOYSA-N acridine-1-sulfonamide Chemical compound C1=CC=C2C=C3C(S(=O)(=O)N)=CC=CC3=NC2=C1 BDQRMEBGHYKVLA-UHFFFAOYSA-N 0.000 description 1
- 125000002252 acyl group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000002421 anti-septic effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000000337 buffer salt Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 235000014633 carbohydrates Nutrition 0.000 description 1
- 230000003197 catalytic effect Effects 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 150000002460 imidazoles Chemical class 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- HWYHZTIRURJOHG-UHFFFAOYSA-N luminol Chemical compound O=C1NNC(=O)C2=C1C(N)=CC=C2 HWYHZTIRURJOHG-UHFFFAOYSA-N 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000002493 microarray Methods 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 229910017604 nitric acid Inorganic materials 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000001259 photo etching Methods 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 229960000856 protein c Drugs 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- GEHJYWRUCIMESM-UHFFFAOYSA-L sodium sulfite Chemical class [Na+].[Na+].[O-]S([O-])=O GEHJYWRUCIMESM-UHFFFAOYSA-L 0.000 description 1
- 235000010265 sodium sulphite Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000003319 supportive effect Effects 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a kind of modularization biological sample analysis chip, belong to technical field of biological.Modularization biological sample analysis chip of the invention is used to support, fixes and integrated analysis module comprising baseplate material, analysis module is integrated with substrate, the biomolecule such as albumen, nucleic acid, polypeptide or organic molecule can be coated with analysis module surface as capture molecule；Analysis chip surface further comprises at least a kind of closing component, non-specific adsorption for avoiding albumen or other biological and organic molecule, hydrophilic substrates can be that baseplate material is hydrophilic in itself or hydrophobic material is modified hydrophilic or is changed into hydrophilic nmature by reagent or sealer treatment, and two processes of hydrophilic nmature and closing can reduce non-specific binding and interference.Modularization biological sample analysis chip of the invention solves the problems, such as that current biological sample analysis chip is high to point sample machine requirement, point sample precision is not high and reaction system is not independent enough.
Description
Technical field
The invention belongs to technical field of biological, more particularly to a kind of modularization biological sample analysis chip.
Background technology
Into 21 century, biochip technology has welcome the development of high speed, automatic printing or light guiding chemical synthesising technology
On the materials such as silicon chip, glass, gel or nylon membrane manufacture micro-array biomolecule chip, realize to compound, protein,
The detection of nucleic acid, cell or other biological components wide coverage.
Be mainly and be coupled for albumen equimolecular or is coated on substrate and is detected by the technology of preparing of current biochip,
Patent 200710134477.9 discloses a kind of coated chip of covalent coupling, and patent US20040009528A1 discloses one kind
The chip of the covalently coating with sulfydryl, and these methods are all direct coateds on the substrate of chip, and it is anti-in detection process
Molecule to be coupled typically all is configured to solution by the package amount of body by a model machine, and point sample is coated with chip, patent
01105795.5 discloses a kind of method that utilization high speed spotting methods prepare biochip, but utilizes the method for point sample to machine
The requirement of device is high, even if best machine also can be faulty or be loaded situation about failing, and such case is difficult detection,
It is simultaneously an independent coupling reaction system between each drop and substrate, patent 200510013108.5 discloses utilization
Photoetching technique is processed to substrate, and retain effective response area has carried out certain improvement to technology, but still solves
The not problem of point sample precision and reaction system independence, each point of sample is a coating system in theory, this is from principle
Application potential of the biochip in quantitative determination is limited, this is also the current main application pain spot of biochip.
The content of the invention
The embodiment of the present invention provides a kind of modularization biological sample analysis chip.The present invention changes traditional point sample thinking,
Invented a kind of modularization high accuracy analysis chip, it is intended to solve current biological sample analysis chip it is high to point sample machine requirement,
Point sample precision is not high and not independent enough the problem of reaction system.
The embodiment of the present invention is achieved in that a kind of modularization biological sample analysis chip, including water-wetted surface base
Plate material, analysis module；The baseplate material of described water-wetted surface is used to support, fixes and integrated analysis module；The analysis
Module assembled is on baseplate material；Capture molecule is coated with described analysis module surface；The analysis module surface and group
Baseplate material surface after dress analysis module comprises at least a kind of closing component, it is described close component for avoid albumen or other
The non-specific adsorption of biological and organic molecule；Described analysis module is to be assembled on baseplate material after the completion of being coated with.
Hydrophilic substrates in the present invention can be for baseplate material is hydrophilic in itself or hydrophobic material is modified hydrophilic or passes through
Reagent or sealer treatment are changed into hydrophilic nmature, and two processes of hydrophilic nmature and closing can reduce non-specific binding and do
Disturb.
Described baseplate material is in the baseplate material of metal material, glass material, silicon materials or high molecule plastic material
One kind.
The water-wetted surface is that baseplate material is hydrophilic in itself, hydrophobic material is modified hydrophilic or is changed into by sealer treatment
The water-wetted surface of hydrophilic nmature.
Described capture molecule is used to capture the material to be analyzed in biological sample, and the capture molecule is from albumen, sugar
Albumen, polypeptide, nucleic acid, organic molecule, polysaccharide or antibody class biomolecule.
Described analysis module has carboxyl, amino, chloromethyl, epoxy alkyl, biotin, aldehyde radical or mercapto selected from surface
The material for possessing conjugated biological molecules ability of base modification；Possesses the metal material of conjugated biological molecules ability；And surface tool
For the one kind in the polystyrene of Biomolecular adsorption ability, nylon or cellulose acetate material.
Described closing molecule is bovine serum albumin(BSA), tween, skimmed milk power, casein, amino acid or xylitol tool
There are the biology or organic molecule of hydrophobic binding characteristic.
Described modularization biological sample analysis chip is further used as module assembled and constitutes new on bigger substrate
Chip.
Described modularization biological sample analysis chip, also including the reference module for internal reference, reference module is directly marked
Note has the analog of determinand or determinand, for carrying out reference to chip and to detection signal.
A kind of biological sample analysis method of the above-mentioned modularization biological sample analysis chip of application, comprises the following steps：
(1) capture molecule is coated in the materials A for possessing adsorption capacity or covalent coupling ability or cuts into material
Small module is coated with；
(2) it is assembled in after being cut by the materials A of well cutting or by materials A on baseplate material B, or materials A is assembled
Cut on substrate B, formed analysis chip C, analysis chip C can further be cut the multiple chips of generation or be analyzed by multiple
Chip C is further assembled in the new analysis chip formed on substrate；
(3) analysis chip C is closed with the reagent containing closing component；
(4) analysis chip C is contacted in molecules detected thing；
(5) cleaning analysis chip C；
(6) analysis chip C with containing being contacted with markd molecular agents；
(7) filled with signal acquisition by light source activation if the mark molecule of step (6) is for fluorescence molecule or quantum dot
Put and detected；If the mark molecule of step (6) be enzyme or direct light emitting molecule, analysis chip further with reaction reagent
Contact is formed luminous signal and is detected using signal pickup assembly.
Described signal pickup assembly can be selected from CCD (charge coupled device), PMT (photomultiplier), PD (photoelectric tube),
It is preferred that CCD.
A kind of kit, including
(1) above-mentioned modularization biological sample analysis chip；
(2) cleaning reagent；
(3) there is the reagent of binding ability component containing markd and checking matter.
(4) if the mark in (3) is or direct light emitting molecule, kit further includes a kind of luminescence-producing reaction
Reagent.
The present invention has the following advantages that compared with prior art：
The present invention changes traditional point sample thinking, has invented a kind of modularization high accuracy analysis chip, it is intended to solve current
Biological sample analysis chip is high to the requirement of point sample machine, the problem that point sample precision is not high and reaction system is not independent enough.The present invention
Modularization biological sample analysis chip be used to supporting comprising baseplate material, fixed and integrated analysis module, be integrated with substrate
Analysis module, can be coated with the biomolecule such as albumen, nucleic acid, polypeptide or organic molecule as capture point on analysis module surface
Son；Analysis chip surface further comprises at least a kind of closing component, for avoiding albumen or other biological and organic molecule
Non-specific adsorption, hydrophilic substrates can for baseplate material is hydrophilic in itself or hydrophobic material is modified hydrophilic or by reagent or
Sealer treatment is changed into hydrophilic nmature, and two processes of hydrophilic nmature and closing can reduce non-specific binding and interference.
Brief description of the drawings
Fig. 1 is coating, cutting, assembling, the closed process exemplary plot of modularization biological sample analysis chip, wherein (a) is first
Coating cuts assembling again, and (b) first cuts and be coated with assembling again, and (c) is cut during assembling, and is cut after (d) assembling
Assemble again.
Fig. 2 is the schematic diagram of modularization biological sample analysis chip.
Fig. 3 is the assembling schematic diagram of analysis module and baseplate material；(a) is wherein schemed for analysis module is assembled to depression
On substrate；Figure (b) is flushed using cover plate covering adjustment analysis module surface with substrate surface.
Specific embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, it is right below in conjunction with drawings and Examples
The present invention is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, and
It is not used in the restriction present invention.
Embodiment 1
The invention provides a kind of modularization biological sample analysis chip, including water-wetted surface baseplate material, analysis mould
Block；The baseplate material of described water-wetted surface is used to support, fixes and integrated analysis module；The analysis module is assembled in substrate
On material；Capture molecule is coated with described analysis module surface；Behind the analysis module surface and Assembly analysis module
Baseplate material surface comprises at least a kind of closing component, and described closes component for avoiding albumen or other biological and organic molecule
Non-specific adsorption；Described analysis module is to be assembled on baseplate material after the completion of being coated with.
Hydrophilic substrates in the present invention can be for baseplate material is hydrophilic in itself or hydrophobic material is modified hydrophilic or passes through
Reagent or sealer treatment are changed into hydrophilic nmature, and two processes of hydrophilic nmature and closing can reduce non-specific binding and do
Disturb.
Described baseplate material is in the baseplate material of metal material, glass material, silicon materials or high molecule plastic material
One kind.
The water-wetted surface is that baseplate material is hydrophilic in itself, hydrophobic material is modified hydrophilic or is changed into by sealer treatment
The water-wetted surface of hydrophilic nmature.
Described capture molecule is used to capture the material to be analyzed in biological sample, and the capture molecule is from albumen, sugar
Albumen, polypeptide, nucleic acid, organic molecule, polysaccharide or antibody class biomolecule.
Described analysis module has carboxyl, amino, chloromethyl, epoxy alkyl, biotin, aldehyde radical or mercapto selected from surface
The material for possessing conjugated biological molecules ability of base modification；Possesses the metal material of conjugated biological molecules ability；And surface tool
For the one kind in the polystyrene of Biomolecular adsorption ability, nylon or cellulose acetate material.
Described closing molecule is bovine serum albumin(BSA), tween, skimmed milk power, casein, amino acid or xylitol tool
There are the biology or organic molecule of hydrophobic binding characteristic.
Described modularization biological sample analysis chip is further used as module assembled and constitutes new on bigger substrate
Chip.
Described modularization biological sample analysis chip, also including the reference module for internal reference, reference module is directly marked
Note has the analog of determinand or determinand, for carrying out reference to chip and to detection signal.
A kind of biological sample analysis method of the above-mentioned modularization biological sample analysis chip of application, comprises the following steps：
(1) capture molecule is coated in the materials A for possessing adsorption capacity or covalent coupling ability or cuts into material
Small module is coated with；
(2) it is assembled in after being cut by the materials A of well cutting or by materials A on baseplate material B, or materials A is assembled
Cut on substrate B, formed analysis chip C, analysis chip C can further be cut the multiple chips of generation or be analyzed by multiple
Chip C is further assembled in the new analysis chip formed on substrate；
(3) analysis chip C is closed with the reagent containing closing component；
(4) analysis chip C is contacted in molecules detected thing；
(5) cleaning analysis chip C；
(6) analysis chip C with containing being contacted with markd molecular agents；
(7) filled with signal acquisition by light source activation if the mark molecule of step (6) is for fluorescence molecule or quantum dot
Put and detected；If the mark molecule of step (6) be enzyme or direct light emitting molecule, analysis chip further with reaction reagent
Contact is formed luminous signal and is detected using signal pickup assembly.
Described signal pickup assembly can be selected from CCD (charge coupled device), PMT (photomultiplier), PD (photoelectric tube),
It is preferred that CCD.
A kind of kit, including
(1) above-mentioned modularization biological sample analysis chip；
(2) cleaning reagent；
(3) there is the reagent of binding ability component containing markd and checking matter.
(4) if the mark in (3) is or direct light emitting molecule, kit further includes a kind of luminescence-producing reaction
Reagent.
A kind of preparation method of modularization biological sample analysis chip, comprises the following steps that：
Capture molecule is coated in first on the material for possessing adsorption capacity or covalent coupling ability, the material can use phase
Material is carried out into cutting to larger area, after coating it is assembled in form analysis chip on baseplate material, analysis chip can
After multiple chip modules or multiple cuttings being produced with further cutting further on assembling and substrate (in including but not limited to Fig. 1
Assembling process shown in (d))；Or cut before coating, then it is coated with, it is assembled on substrate after coating；Or bag
By it after further being cut after substrate mounting；In including but not limited to Fig. 1 (a), (b), (c), the chip manufacturing mistake shown in (d)
Journey.
Modularization chip of the invention as shown in Figure 2 is assembled by substrate, analysis module, and baseplate material includes but do not limit
Possess the material of supportive in metal, glass, nylon, silicon, high molecule plastic etc., multiple analysis modules can be assembled on substrate,
Also (d) is shown during multiple multiple modules such as Fig. 1 for turning into unit by analysis module and substrate in batch can be assembled, analysis module surface bag
There is biomolecule, biomolecule includes but is not limited to biological point of albumen, antibody, polypeptide, amino acid, carbohydrate, enzyme, nucleic acid etc.
Son and organic molecule, further comprising a kind of closing molecule, closing molecule is used to close the non-specific of chip surface chip surface
Property binding site, closing molecule include be not limited to BSA, amino acid, sugar, milk, casein, surfactant, polyethylene glycol, sugar
The biomolecule and organic molecule with hydrophobic binding characteristic such as alcohol, the analysis module of different chip is by same reaction condition
Generation, substrate can be fabricated to sunk structure, and chip is placed on it and holding surface smooth as shown in Figure 3.
The analysis method of the biological specimen of present invention invention：(1) capture molecule is coated in and possesses adsorption capacity or covalent
In the materials A of coupling capacity or material is cut into small module to be coated with；(2) enter by the materials A of well cutting or by materials A
It is assembled on baseplate material B after row cutting, or materials A is assembled on substrate B is cut, form analysis chip C, analyzes core
Piece C can further cut many analysis chips of generation, and the chip C after cutting can further be assembled in the new of formation on substrate and divide
Analysis chip；(2) analysis chip C is closed with the reagent containing closing component；(3) by analysis chip C in molecules detected thing
Contact；(4) cleaning analysis chip C；(5) analysis chip C with containing being contacted with markd molecular agents；(6) if the mark of step 6
Son score for fluorescence molecule or quantum dot then utilize light source activation, is detected using fluorescence detector；(7) if step 6
Mark molecule is enzyme or direct light emitting molecule, and analysis chip further contacts to form luminous signal using luminous with reaction reagent
Signal detector is detected；The closing of analysis chip can be to fit together, also after substrate and analysis module individually closing
Can analysis module together with substrate mounting after closed, the molecule of mark include but is not limited to fluorescence molecule, quantum dot, can
The luminous enzyme of catalytic substrate, molecule that can be directly luminous by reacting triggering.It is of the invention in the detection process of signal Analysis
In method, need to be excited using light source if the label of mark molecule is for fluorescence molecule or quantum dot, recycle letter
Number collection CCD or PMT or PD (photoelectric tube) carry out signal acquisition, if mark molecule for can trigger chemiluminescent point
Son, then first carry out luminous triggering reaction, and such as horseradish peroxidase-labeled needs to be reacted with luminol substrate reagent
To trigger chemiluminescence signal, chip substrate of the invention can also be integrated with telegraph circuit for reacting generation telecommunications on chip
Number detection.
Chip of the invention can make a kind of kit, in kit include chip, included on the chip substrate and
Integrated analysis module, analysis module is coated with capture molecule, on chip containing closing molecule can effectively reduce it is non-specific
Property combination, comprising cleaning reagent in kit, cleaning reagent comprises at least a kind of buffer salt and a kind of antiseptic ingredient and uses
In the non-specific binding molecule and luminous substrate molecule of cleaning chip, labelled reagent is included in kit, in the reagent
Mark molecule can be combined with captured molecule, and capture molecule, captured molecule, mark molecule form complex.If mark
Molecule is the inspection that fluorescence molecule or quantum dot molecule can directly be carried out by optical excitation and CCD or PMT or PD (photoelectric tube)
Survey.If the label of mark molecule is enzyme, kit is further included if horseradish peroxidase or alkaline phosphatase
A kind of substrate reagent, contains luminous substrate in substrate reagent, if the label of mark molecule is direct light emitting molecule, such as acridine
Ester, acridine sulfonamide, acridine acyl ammonia or its analog, then react shape containing reaction reagent in substrate reagent with direct light emitting molecule
Into luminous signal, signal detection is carried out by CCD or PMT.
Application Example：
Reagent 1:
Tris 10mL 0.01M/L；HCL 0.0003N/L；
Regulation pH to 8.5,0.22 μm filters are filtered
Reagent 2:
Imidazoles 2.72g/L；Tween 0.2mL/l；NaCL 8.76g/L；8.5,0.22 μm of pH to, filter is filtered
Reagent 3:
0.58g/L Na2HPO4,0.05g/L KCl, 0.05g/L K3PO4,2.0g/L NaCl, BSA 10g/L, TWEEN
4mL/l；PH 7.4., 0.22 μm of filter is filtered
Reagent 4:
Mab PCT 16B5 Cat.#4PC47 hytest 1mg/mL
Sodium azide 0.1%
Reagent 5:
Mab PCT 42 Cat.#4C10 hytest 2mg/mL
Sodium azide 0.1%
Reagent 6：
PCT recombinant protein c at.#8PC5 50ng/mL
Sodium azide 0.1%
Reagent 7:
Mab CRP C2 Cat.#4C28 hytest 1mg/mL
Sodium azide 0.1%
Reagent 8：
Mab CRP C6 Cat.#4C28 hytest 2mg/mL
Sodium azide 0.1%
Reagent 9
The anti-source Cat.#8C72 hytest 20ug/mL of CRP
Sodium azide 0.1%
Reagent 10
HNO30.15M；H2O20.7%, Triton-100 0.25%；NaOH 0.125M；
Reagent 11
9- (4- chlorophenylthio benzoyls Oxymethylene) -10- methyl -9,10- acridans-disodium salt 100mg/L, N,
N dimethyl acridine nitrate, 1.0mg/L lauryl sodium sulfate, 0.8g/L sodium sulfites, 5mg/L TWEEN-20,0.31g/L
The Tris buffer systems PH 9.10 of 0.26mol/L.
Application Example 1
The Polystyrene plastic plate cleaning, drying of reagent 1 of 100mmX100mmX1mm is taken, is positioned in plate, by reagent 4
Protein concentration is diluted to 20ug/mL, adds and submerge in plate XPS, room temperature 2 hours, and 4 DEG C are spent night；With reagent to flat
Plate is cleaned, and is dried under cleaning condition.The fritter (hereinafter referred to as PCT analysis modules) of precision laser cutting 5mmX5mm；
The Polystyrene plastic plate cleaning, drying of reagent 1 of 100mmX100mmX1mm is taken, is positioned in plate, by reagent 7
Protein concentration is diluted to 15ug/mL, adds and submerge in plate XPS, room temperature 2 hours, and 4 spend night；With reagent to flat board
Cleaned, and dried under cleaning condition.The fritter (hereinafter referred to as CRP analysis modules) of precision laser cutting 5mmX5mm；
Cutting glass plate is into 1cmX3cmx2mm fritters as substrate；PCT analysis modules and CRP points are adhered on each substrate
Analysis module, forms analysis chip；
Analysis chip is placed and closed as injected reagent 3 in plate, and room temperature is taken out for 3 hours；
Take reagent 580ul and add 600ul reagents 1, be subsequently adding the 0.5mmol/l acridinium esters of 150ul, mix, room temperature is kept away
Light 20 minutes, adds lysine, 180ul reagents 1 to place 30 minutes, obtains acridinium ester label.PCT- labelled reagents,
Take reagent 870ul and add 600ul reagents 1, be subsequently adding the 0.5mmol/l acridinium esters of 150ul, mix, room temperature is kept away
Light 20 minutes, adds lysine, 180ul reagents 1 to place 30 minutes, obtains acridinium ester label.CRP- labelled reagents；
By PCT- labelled reagents and CRP labelled reagents according to 1：1 ratio is mixed, and obtains PCT-CRP labelled reagents；
Reagent 6 is diluted into PCT concentration, the dilution configuration of reagent 9：
Standard liquid 1：PCT 0.1ng/mL CRP 0.5ug/mL；Standard liquid 2：PCT 0.5ng/mL CRP 1ug/
mL；Standard liquid 3：PCT 1ng/mL CRP 2ug/mL；Standard liquid 4：PCT 10ng/mL CRP 10ug/mL；Standard liquid
5：PCT 30ng/mL CRP 20ug/mL；
Testing process is as follows, and (as sample) in standard liquid is immersed in analysis chip, is incubated at room temperature 5 minutes, with examination
The cleaning of agent 2 chip twice, then analysis chip is immersed in PCT-CRP labelled reagents is incubated 5 minutes, is cleaned with cleaning reagent 2
Chip twice, chip is placed in reagent 10 while carrying out CCD detection luminous signals.
After standard reagent is detected and sets up luminous signal standard curve, with reagent 6 and the addition serum of reagent 9, pass through
Weight method configuration is obtained, sample 1：CRP 2ug/MLPCT 0.5ng/mL, sample 2：CRP 5ug/MLPCT 5ng/mL, sample 3：
CRP 10ug/MLPCT 30ng/mL, repeat above-mentioned flow sample is tested with analysis chips, the test result such as institute of table 1
Show：
Table 1
| CRP(ug/ml) | Configuration concentration | Chip 1 | Chip 2 | Chip 3 | Chip 4 | Chip 5 | Chip 6 | Chip 7 | Chip 8 | CV | Average |
| Sample 1 | 2 | 2.10 | 2.10 | 2.04 | 2.10 | 2.20 | 1.90 | 2.20 | 2.20 | 4.86% | 2.11 |
| Sample 2 | 5 | 5.15 | 5.15 | 5.15 | 5.15 | 4.85 | 4.85 | 4.85 | 5.15 | 3.08% | 5.04 |
| Sample 3 | 10 | 10.80 | 10.40 | 9.40 | 9.60 | 9.60 | 9.80 | 9.90 | 10.40 | 4.92% | 9.99 |
| PCT(ng/ml) | Configuration concentration | Chip 1 | Chip 2 | Chip 3 | Chip 4 | Chip 5 | Chip 6 | Chip 7 | Chip 8 | CV | Average |
| Sample 1 | 0.5 | 0.51 | 0.48 | 0.52 | 0.52 | 0.53 | 0.56 | 0.53 | 0.48 | 5.45% | 0.51 |
| Sample 2 | 5 | 5.15 | 5.15 | 5.15 | 5.00 | 5.15 | 4.90 | 5.15 | 5.80 | 5.16% | 5.18 |
| Sample 3 | 30 | 30.90 | 29.10 | 29.10 | 30.90 | 29.10 | 29.10 | 29.10 | 29.10 | 2.82% | 29.55 |
Application Example 2
Epoxy alkyl modification chip (telecheminternational CAT：SME2 25X76mm) carried out clearly with reagent 1
Wash, be positioned in plate, the protein concentration of reagent 4 is diluted to 20ug/mL, XPS is submerged in addition plate, room temperature 2 is small
When, 4 spend night；Flat board is cleaned with reagent, and is dried under cleaning condition.The fritter of precision laser cutting 5mmX5mm
(hereinafter referred to as PCT analysis modules)；
Epoxy alkyl modification chip (telecheminternational CAT：SME2 25X76mm) carried out clearly with reagent 1
Wash, be positioned in plate, the protein concentration of reagent 7 is diluted to 15ug/mL, XPS is submerged in addition plate, room temperature 2 is small
When, 4 spend night；Flat board is cleaned with reagent, and is dried under cleaning condition.The fritter of precision laser cutting 5mmX5mm
(hereinafter referred to as PCT analysis modules)；
Cutting glass plate is into 1cmX3cmx2mm fritters as substrate；PCT analysis modules and CRP points are adhered on each substrate
Analysis module, forms analysis chip；
Reagent 5,300uL is taken, 14uL Sulfo-NHS-LC-Biotinylation Kit (THERMO) are added, 37 DEG C incubate
Educate 39 minutes, add in super filter tube, 12000Xg is centrifuged 10 minutes, and pipettor is mixed and is centrifuged again 10 minutes, add 0.3mL marks
Buffer solution 6000Xg is centrifuged 10 minutes.The solution in centrifuge tube is collected, supplemental markers buffer solution to 0.5mL adds 0.4mL strepto-s
Avidin marks alkaline phosphatase (THERMO AP Streptavidin 434322) to be incubated at room temperature 30 minutes.By cross-linking agent mistake
Sephadex g200 are purified, and obtain PCT- labelled reagents.
Reagent 8,500uL is taken, 14uL Sulfo-NHS-LC-Biotinylation Kit (THERMO) are added, 37 DEG C incubate
Educate 39 minutes, add in super filter tube, 12000Xg is centrifuged 10 minutes, and pipettor is mixed and is centrifuged again 10 minutes, add 0.3mL marks
Buffer solution 6000Xg is centrifuged 10 minutes.The solution in centrifuge tube is collected, supplemental markers buffer solution to 0.5mL adds 0.6mL strepto-s
Avidin marks alkaline phosphatase (THERMO AP Streptavidin 434322) to be incubated at room temperature 30 minutes.By cross-linking agent mistake
Sephadex g200 are purified, and obtain CRP- labelled reagents.
PCT labelled reagents are mixed in proportion with CRP labelled reagents 1: 1；
Analysis chip is placed and closed as injected reagent 3 in plate, and room temperature is taken out for 3 hours；
Reagent 6 is diluted into PCT concentration, the dilution configuration of reagent 9：
Standard liquid 1：CRP 0.5ug/mL；Standard liquid 2：PCT 0.5ng/mL CRP 1ug/mL；Standard liquid 3：
PCT 1ng/mL CRP 2ug/mL；Standard liquid 4：PCT 10ng/mL CRP 10ug/mL；Standard liquid 5：PCT 30ng/mL
CRP 20ug/mL：
Testing process is as follows, and (as sample) in standard liquid is immersed in analysis chip, is incubated at room temperature 5 minutes, with examination
The cleaning of agent 2 chip twice, then analysis chip is immersed in PCT-CRP labelled reagents is incubated 5 minutes, is cleaned with cleaning reagent 2
Chip twice, chip is placed in reagent 11 while carrying out CCD detection luminous signals.
After standard reagent is detected and sets up luminous signal standard curve, with reagent 6 and the addition serum of reagent 9, pass through
Weight method configuration is obtained, sample 1：CRP 2ug/MLPCT 0.5ng/mL, sample 2：CRP 5ug/mLPCT 5ng/mL, sample 3：
CRP 10ug/MLPCT 30ng/mL, repeat above-mentioned flow sample is tested with analysis chips, the test result such as institute of table 2
Show：
Table 2
| CRP(ug/ml) | Configuration concentration | Chip 1 | Chip 2 | Chip 3 | Chip 4 | Chip 5 | Chip 6 | Chip 7 | Chip 8 | CV | Average |
| Sample 1 | 2 | 1.92 | 1.92 | 1.92 | 2.08 | 2.08 | 2.08 | 2.08 | 1.92 | 4.28% | 2.00 |
| Sample 2 | 5 | 4.85 | 5.15 | 4.85 | 5.15 | 4.85 | 4.85 | 4.85 | 5.15 | 3.13% | 4.96 |
| Sample 3 | 10 | 10.25 | 9.75 | 9.75 | 9.75 | 9.75 | 10.25 | 9.75 | 9.75 | 2.34% | 9.88 |
| PCT(ng/ml) | Configuration concentration | Chip 1 | Chip 2 | Chip 3 | Chip 4 | Chip 5 | Chip 6 | Chip 7 | Chip 8 | CV | Average |
| Sample 1 | 0.5 | 0.53 | 0.48 | 0.48 | 0.53 | 0.53 | 0.48 | 0.53 | 0.48 | 5.35% | 0.50 |
| Sample 2 | 5 | 5.15 | 4.85 | 5.15 | 5.15 | 5.15 | 5.15 | 5.15 | 4.85 | 2.74% | 5.08 |
| Sample 3 | 30 | 30.81 | 30.81 | 30.81 | 30.81 | 30.81 | 29.19 | 29.19 | 29.19 | 2.78% | 30.20 |
Application Example 3
Epoxy alkyl modification chip (telecheminternational CAT：SME2 25X76mm) carried out clearly with reagent 1
Wash, be positioned in plate, the protein concentration of reagent 4 is diluted to 20ug/mL, XPS is submerged in addition plate, room temperature 2 is small
When, 4 spend night；Flat board is cleaned with reagent, and is dried under cleaning condition.The fritter of precision laser cutting 5mmX5mm
(hereinafter referred to as PCT analysis modules)；
Epoxy alkyl modification chip (telecheminternational CAT：SME2 25X76mm) carried out clearly with reagent 1
Wash, be positioned in plate, the protein concentration of reagent 7 is diluted to 10ug/mL, XPS is submerged in addition plate, room temperature 2 is small
When, 4 spend night；Flat board is cleaned with reagent, and is dried under cleaning condition.The fritter of precision laser cutting 5mmX5mm
(hereinafter referred to as PCT analysis modules)；
Cutting glass plate is into 1cmX3cmx2mm fritters as substrate；PCT analysis modules and CRP points are adhered on each substrate
Analysis module, forms analysis chip；
Analysis chip is placed and closed as injected reagent 3 in plate, and room temperature is taken out for 3 hours；
By fluorescent latex microspherulite diameter 100nm, excitation wavelength 480nm pH 6.0,50mM the MES buffering with carboxyl
Liquid is cleaned, and diluted concentration is 0.01mg/mL, takes 2mL, adds EDC (5mg/mL) 200uL, is mixed and is added sulfo-NHS
(2mg/ML) 200uL, reaction priming reaction 0.5 hour, reagent 5 adds 80uL room temperature reactions 2 hours, adds the second of 0.01g/mL
Diamines 500uL, reacts 1 hour, and centrifugation buffer solution for cleaning adds reagent 3 to clean twice, adds buffering agents 3,10mL, obtains
Obtain PCT- microballoon labelled reagents.
By fluorescent latex microspherulite diameter 100nm, excitation wavelength 480nm pH 6.0,50mM the MES buffering with carboxyl
Liquid is cleaned, and diluted concentration is 0.01mg/mL, takes 2mL, adds EDC (5mg/mL) 200uL, is mixed and is added sulfo-NHS
(2mg/ML) 200uL, reaction priming reaction 0.5 hour, reagent 8 adds 60uL room temperature reactions 2 hours, adds the second of 0.01g/mL
Diamines 500uL, reacts 1 hour, and centrifugation buffer solution for cleaning adds reagent 3 to clean twice, adds buffering agents 3,10mL, obtains
Obtain CRP microballoon labelled reagents.
PCT- microballoons mark is related to carry out mixing acquisition PCT-CRP microballoon labelled reagents with CRP microballoons labelled reagent；
Reagent 6 is diluted into PCT concentration, the dilution configuration of reagent 9；
Standard liquid 1：CRP 0.5ug/mL；Standard liquid 2：PCT 0.5ng/mL CRP 1ug/mL；Standard liquid 3：
PCT 1ng/mL CRP 2ug/mL；Standard liquid 4：PCT 10ng/mL CRP 10ug/mL；Standard liquid 5：PCT 30ng/mL
CRP 20ug/mL；
Testing process is as follows, and (as sample) in standard liquid is immersed in analysis chip, is incubated at room temperature 5 minutes, with examination
The cleaning of agent 2 chip twice, then analysis chip is immersed in PCT-CRP labelled reagents is incubated 5 minutes, is cleaned with cleaning reagent 2
Chip twice, with laser excitation, while detecting fluorescence signal using PMT.
After standard reagent is detected and sets up luminous signal standard curve, with reagent 6 and the addition serum of reagent 9, pass through
Weight method configuration is obtained, sample 1：CRP 2ug/MLPCT 0.5ng/mL, sample 2：CRP 5ug/MLPCT 5ng/mL, sample 3：
CRP 10ug/MLPCT 30ng/mL, repeat above-mentioned flow sample is tested with analysis chips, test result such as table 3：
Table 3
| cRP(ug/ml) | Configuration concentration | Chip 1 | Chip 2 | Chip 3 | Chip 4 | Chip 5 | Chip 6 | Chip 7 | Chip 8 | CV | Average |
| Sample 1 | 2 | 1.86 | 2.10 | 2.10 | 2.14 | 1.90 | 1.86 | 1.86 | 1.86 | 6.55% | 1.96 |
| Sample 2 | 5 | 5.30 | 5.30 | 5.20 | 5.30 | 5.10 | 4.70 | 5.30 | 5.10 | 4.70 | 5.16 |
| Sample 3 | 10 | 9.40 | 9.40 | 10.60 | 10.60 | 9.40 | 10.50 | 10.50 | 9.40 | 6.17% | 9.98 |
| PcT(ng/ml) | Configuration concentration | Chip 1 | Chip 2 | Chip 3 | Chip 4 | Chip 5 | Chip 6 | Chip 7 | Chip 8 | CV | Average |
| Sample 1 | 0.5 | 0.46 | 0.46 | 0.46 | 0.49 | 0.47 | 0.54 | 0.46 | 0.46 | 5.95% | 0.48 |
| Sample 2 | 5 | 4.65 | 4.65 | 4.65 | 5.35 | 4.65 | 5.35 | 4.65 | 4.65 | 6.72% | 4.83 |
| Sample 3 | 30 | 32.10 | 27.90 | 27.90 | 27.90 | 27.90 | 32.10 | 32.10 | 27.90 | 7.37% | 29.48 |
Presently preferred embodiments of the present invention is the foregoing is only, is not intended to limit the invention, it is all in essence of the invention
Any modification, equivalent and improvement made within god and principle etc., should be included within the scope of the present invention.
Claims (10)
1. a kind of modularization biological sample analysis chip, it is characterised in that：Baseplate material, analysis module including water-wetted surface；
The baseplate material of described water-wetted surface is used to support, fixes and integrated analysis module；The analysis module is assembled in substrate material
On material；Capture molecule is coated with described analysis module surface；Base behind the analysis module surface and Assembly analysis module
Plate material surface comprises at least a kind of closing component, and described component of closing is for avoiding albumen or other biological and organic molecule
Non-specific adsorption；Described analysis module is to be assembled on baseplate material after the completion of being coated with.
2. modularization biological sample analysis chip as claimed in claim 1, it is characterised in that：Described baseplate material is metal
One kind in the baseplate material of material, glass material, silicon materials or high molecule plastic material.
3. modularization biological sample analysis chip as claimed in claim 1, it is characterised in that：The water-wetted surface is substrate material
Material is hydrophilic in itself, hydrophobic material is modified hydrophilic or the water-wetted surface for being changed into hydrophilic nmature is processed by sealer.
4. modularization biological sample analysis chip as claimed in claim 1, it is characterised in that：Described capture molecule is used to catch
Obtain the material to be analyzed in biological sample, the capture molecule is from albumen, glycoprotein, polypeptide, nucleic acid, organic molecule, many
Sugar or antibody class biomolecule.
5. modularization biological sample analysis chip as claimed in claim 1, it is characterised in that：Described analysis module is selected from
What surface had carboxyl, amino, chloromethyl, epoxy alkyl, biotin, aldehyde radical or a sulfydryl modification possesses conjugated biological molecules ability
Material；Possesses the metal material of conjugated biological molecules ability；And surface possess Biomolecular adsorption ability polystyrene,
One kind in nylon or cellulose acetate material.
6. modularization biological sample analysis chip as claimed in claim 1, it is characterised in that：Described closing molecule is ox blood
Pure albumen, tween, skimmed milk power, casein, amino acid or xylitol have biological or organic point of hydrophobic binding characteristic
Son.
7. the modularization biological sample analysis chip described in claim 1 is further used as module assembled group on bigger substrate
The chip of Cheng Xin.
8. modularization biological sample analysis chip as claimed in claim 1, it is characterised in that：Also include the reference for internal reference
Module, reference module is directly labeled with the analog of determinand or determinand, for carrying out reference to chip and to detection signal.
9. a kind of biological sample analysis method of modularization biological sample analysis chip described in application claim 1, its feature exists
In：Comprise the following steps：
(1) capture molecule is coated in the materials A for possessing adsorption capacity or covalent coupling ability or is cut into material small
Module is coated with；
(2) it is assembled in after being cut by the materials A of well cutting or by materials A on baseplate material B, or materials A is assembled in base
Cut on plate B, formed analysis chip C, analysis chip C can further be cut the multiple chips of generation or by multiple analysis chips
C is further assembled in the new analysis chip formed on substrate；
(3) analysis chip C is closed with the reagent containing closing component；
(4) analysis chip C is contacted in molecules detected thing；
(5) cleaning analysis chip C；
(6) analysis chip C with containing being contacted with markd molecular agents；
(7) entered with signal pickup assembly by light source activation if the mark molecule of step (6) is for fluorescence molecule or quantum dot
Row detection；If the mark molecule of step (6) is enzyme or direct light emitting molecule, analysis chip is further contacted with reaction reagent
Luminous signal is formed to be detected using signal pickup assembly.
10. a kind of kit, it is characterised in that：Including
(1) analysis chip described in claim 1；
(2) cleaning reagent；
(3) there is the reagent of binding ability component containing markd and checking matter.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710023509.1A CN106872425B (en) | 2017-01-13 | 2017-01-13 | Modularized biological sample analysis chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710023509.1A CN106872425B (en) | 2017-01-13 | 2017-01-13 | Modularized biological sample analysis chip |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106872425A true CN106872425A (en) | 2017-06-20 |
| CN106872425B CN106872425B (en) | 2020-02-18 |
Family
ID=59157556
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710023509.1A Active CN106872425B (en) | 2017-01-13 | 2017-01-13 | Modularized biological sample analysis chip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106872425B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112326949A (en) * | 2017-11-10 | 2021-02-05 | 深圳市真迈生物科技有限公司 | Surface chemical modification method, chip preparation method and chip |
| CN115993350A (en) * | 2021-10-18 | 2023-04-21 | 中国科学院化学研究所 | A fully printed biological detection chip and its preparation method and application |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1204363A (en) * | 1995-10-06 | 1999-01-06 | 麦克罗克隆宁Cccd公司 | Compact cell culture slide |
| CN1472339A (en) * | 2002-08-02 | 2004-02-04 | � 赵 | High-flux cell biological chip testing technology and reagent case |
| CN1641355A (en) * | 2004-12-20 | 2005-07-20 | 山东省医药生物技术研究中心 | Protein chip for detecting blood and cerebro spinal fluid pathogen antibody, and its preparing method and use |
| CN101315369A (en) * | 2008-07-08 | 2008-12-03 | 东南大学 | Multiplexed immunoassays in hydrogels complex-encoded in color and shape |
| CN203535049U (en) * | 2013-11-04 | 2014-04-09 | 内蒙古民族大学 | Kit for quickly detecting salmonella pullorum antibody |
-
2017
- 2017-01-13 CN CN201710023509.1A patent/CN106872425B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1204363A (en) * | 1995-10-06 | 1999-01-06 | 麦克罗克隆宁Cccd公司 | Compact cell culture slide |
| CN1472339A (en) * | 2002-08-02 | 2004-02-04 | � 赵 | High-flux cell biological chip testing technology and reagent case |
| CN1641355A (en) * | 2004-12-20 | 2005-07-20 | 山东省医药生物技术研究中心 | Protein chip for detecting blood and cerebro spinal fluid pathogen antibody, and its preparing method and use |
| CN101315369A (en) * | 2008-07-08 | 2008-12-03 | 东南大学 | Multiplexed immunoassays in hydrogels complex-encoded in color and shape |
| CN203535049U (en) * | 2013-11-04 | 2014-04-09 | 内蒙古民族大学 | Kit for quickly detecting salmonella pullorum antibody |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112326949A (en) * | 2017-11-10 | 2021-02-05 | 深圳市真迈生物科技有限公司 | Surface chemical modification method, chip preparation method and chip |
| CN115993350A (en) * | 2021-10-18 | 2023-04-21 | 中国科学院化学研究所 | A fully printed biological detection chip and its preparation method and application |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106872425B (en) | 2020-02-18 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US6498010B1 (en) | Method for making a device for the simultaneous detection of multiple analytes | |
| EP0874242B1 (en) | Device and apparatus for the simultaneous detection of multiple analytes | |
| CN100386627C (en) | filter protein chip | |
| US20020168699A1 (en) | Rapid classification of biological components | |
| US20040002121A1 (en) | High throughput methods and devices for assaying analytes in a fluid sample | |
| TW201132981A (en) | Enzyme assays on a droplet actuator | |
| KR20000071894A (en) | Multipurpose diagnostic systems using protein chips | |
| JP2004516489A (en) | Protein chip, method for producing the same, detection system using the protein chip, and method using the detection system | |
| CN110849694B (en) | Tacrolimus whole blood sample pretreatment liquid and use method and application thereof | |
| JP2010522885A (en) | Improved antibody profiling sensitivity by increasing reporter antibody stacking | |
| CN117030991A (en) | Extracellular vesicle detection device and detection method | |
| EP2062055B1 (en) | Blood typing | |
| CN106872425A (en) | A kind of modularization biological sample analysis chip | |
| CN106461652A (en) | Control marker for implementing analysis methods on spots | |
| JP5248264B2 (en) | High-sensitivity measurement kit by immunochromatography | |
| RU2298796C2 (en) | Diagnostic system containing protein chips having features for concurrently determining a plurality of indices | |
| JP4197279B2 (en) | Biologically-derived substance detection substrate and manufacturing method thereof | |
| CN1482460A (en) | Immunological measurement method, test piece used, and immunological measurement device | |
| US20150369802A1 (en) | Biomolecule Binding Composite Surfaces, Methods Of Making Such Surfaces, Devices Incorporating Such Surfaces, And Methods Of Using Such Surfaces In Biomolecule Binding Assays, And Devices Therefor | |
| CN119346194B (en) | A high-throughput rapid screening chip for allergens based on 3D microarray and reciprocating flow and its application | |
| CN113125697A (en) | Testosterone homogeneous phase chemiluminescence detection kit and application thereof | |
| KR100785655B1 (en) | Self-assembled monolayer prepared using aminocalixarene derivatives and protein chip manufactured using the self-assembled monolayer | |
| CN121164626A (en) | HBP detection test strip and preparation method thereof | |
| JP2019521367A (en) | Lateral flow sample detection | |
| CN116430035A (en) | Immunochromatography kit with signal capable of being secondarily amplified, preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20171130 Address after: 518000 Guangdong Shenzhen City, Shenzhen Nanshan District Taoyuan Street Li Shan Road University City Pioneer Park 1109 room Applicant after: Shenzhen handed down Biological Medicine Co., Ltd. Address before: 550081 Guanshan Lake District, Guizhou Province green Liansheng International No. 6,7 Building (6) Unit 1 19-layer 4 room Applicant before: Guizhou root Medical Instrument Co., Ltd. |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |