CN1458525A - Integrated capillary biological chip and its producing method - Google Patents
Integrated capillary biological chip and its producing method Download PDFInfo
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- CN1458525A CN1458525A CN 03141391 CN03141391A CN1458525A CN 1458525 A CN1458525 A CN 1458525A CN 03141391 CN03141391 CN 03141391 CN 03141391 A CN03141391 A CN 03141391A CN 1458525 A CN1458525 A CN 1458525A
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Abstract
The integrated capillary biochip is used in the parallel detection, distinction and identification of gene, protein, medicine or acceptor in tested sample. It consists of capillary biochip units mainly, and features that capillary carrier with biological probe is set inside capillary cavity to constitute the unit array for the capillary biochip and that the capillary biochip unit array is cut in perpendicular or non-perpendicular direction to constitute the capillary biochip. The present invention has the advantages of simple structure, high probe density, high sensitivity, high accuracy and no cross talk, and may be produced in large scale.
Description
Technical field
The invention belongs to the life science technical field, more precisely, relate to a kind of integrated capillary biochip that gene, protein (antigen or antibody), medicine or acceptor, tissue or cell, polysaccharide or agglutinin are carried out parallel detection, identification, evaluation.The invention still further relates to the making detection method and the purposes of this integrated capillary biochip.
Background technology
The prior biological chip is meant high, medium and low density nucleic acid (cDNA, mRNA, peptide nucleic acid) probe, protein (polypeptide, enzyme molecule, antigen, antibody), medicine or the acceptor that is coated on the solid phase carrier, the microarray (Microarray) of cell or tissue; Be to utilize the ability that has special mutual identification between biomacromolecule and with their orderly being arranged on the solid phase carrier substrate (film, glass sheet, silicon chip, ceramics etc.), carry out specific reaction or hybridization simultaneously with sample to be checked and corresponding mark biomolecule, can obtain a large amount of useful life science information by automatic reading equipment.
Genetic chip claims the DNA chip again, is a kind of nucleotide (cDNA, mRNA, peptide nucleic acid) probe microarray of high, middle density.Its preparation method mainly contains: the first adopts photoetch and former bit pattern synthesising chemical technology, is solidificated in the nucleic acid probe of a large amount of particular sequences on the carrier substrate in an orderly manner; Its two be with purifying is good the automatic point sample equipment of nucleic acid probe utilization point sample (spray printing) on carrier substrate; It three is to utilize the photoinduction technology that gene probe is grown at a fibre-optic end; It four is to adopt molecular seal method multi-impression original position fixed point DNA to synthesize on carrier substrate; Thereby constitute the genetic chip that stores a large amount of life-information dot matrix.
Protein-chip claims peptide chips again, be that protein is comprised that polypeptide, enzyme molecule, antigen or antibody are fixed on the solid phase carrier substrate, utilize protein molecule can specificity and ligand molecular comprise antigen or antibody, acceptor principle of combining, by corresponding labeled molecule, be used for detecting on a large scale antigen, antibody, acceptor or enzyme molecule, polypeptide fragment then.Its method for making be with purifying is good protein molecule by different mode point samples (spray printing) on solid phase carrier; It two is to adopt photoconduction to peptide synthesis technology, will have specific amino acids polypeptide of sequence chain and be solidificated on the carrier substrate in an orderly manner.
The solid phase carrier substrate that biochip is commonly used has: organic and inorganic polymer film, glass sheet, silicon chip, ceramics etc., such carrier substrate is handled through modifying usually, promptly use various activating reagent by chemical reaction various reactive groups on the carrier substrate surface bond, so that with the biomolecule covalent bond in the point sample thing, formation has the affinity type carrier substrate of different biologic specificities, it is biochip, be used for fixing and detect various active biomolecule, as protein, polypeptide, enzyme, nucleic acid, antigen, antibody, medicine or acceptor, tissue or cell etc., even the various microstructures of structure comprise little hole on carrier substrate, little ditch, microflute, microtubule, microstructure or devices such as Micropump are beneficial to the processing of sample and the distribution of reagent.
Useful light transmitting fiber adopts diverse ways to make the microarray of various biomolecule as solid support material person on fibre-optic top in addition.Capillary fiber or optical fiber have structure trickle (its external diameter only is 5-20 μ m), are easy to the advantage of mass disposal and processing as the carrier of biochip, can fix a large amount of bioprobes and comprise nucleotide probe, enzyme molecule or polypeptide, antigen or antibody, medicine or acceptor, polysaccharide or agglutinin, cell or tissue etc. on the 3-D solid structure of capillary fiber or optical fiber, its probe density can reach ten thousand dot matrix of every square centimeter of 10-400.
Also useful kapillary promptly make the microarray of biomolecule at lumen wall capillaceous, but this scheme has bigger manufacture difficulty as solid support material person.
Authorize Chinese patent 9812010.4 to disclose a kind of " optical fibre matrix type gene chip and preparation method thereof ", its method for making is in advance at the end face growth hormone gene probe of simple optical fiber unit and after solidifying, and the fiber unit with one group of band different genes probe is arranged into array again.The fiber unit direct neighbor that so will carry gene probe is arranged into array, and this technical scheme can not really be avoided the crosstalk phenomenon between adjacent gene probe.
Authorize Chinese utility model patent 01227862.9 to disclose a kind of " kapillary biochip device ", form by the interior transparent capillary of establishing capillary fiber silk (bar), go up point sample object point line-transect (point), respective markers thing, the positive and the negative control line (point) that point has stabilization at capillary fiber silk (bar).This technical scheme only is a small throughput kapillary biochip unit, rather than high flux integrated capillary biochip array, and bioprobe is line (point) shape, but not coating, contaminate, the continuous distribution of growth.
In view of original position is synthesized higher to the host material requirement condition; Directly during point sample owing to adopt complicated technology; regular meeting causes irreversible sex change of generation such as active substance such as DNA or RNA, antigen or antibody, medicine or acceptor, tissue or cell or active loss in the sampling liquid; influence the detection sensitivity and the specificity of biochip, even influenced the stability and the storage life of biochip.Moreover along with increasing of high-density biochip probe density, the rate of dying mutually between adjacent bioprobe increases, and the crosstalk phenomenon between adjacent probe is obviously strengthened, and causes the stability of biochip and accuracy obviously to descend.In addition, because the manufacturing process complexity, respective detection means requirement condition is also very high, causes manufacturing and application cost higher, has limited the wide popularization and application of this new and high technology.
Goal of the invention
Purpose of the present invention is exactly at defective that exists in the prior art or deficiency, especially at the high-density biochip complicated process of preparation with cause the difficult problem of poor stability because of crosstalk phenomenon, provide a kind of probe density high and sensitive, accurately, really do not have crosstalk phenomenon and preparation technology is easy, can carry out the integrated capillary biochip of large-scale industrial production.
Another object of the present invention just provides the method for making of this integrated capillary biochip.
Another object of the present invention just provides this integrated capillary bio-chip test method.
Another object of the present invention just provides the purposes of this integrated capillary biochip.Technical scheme
The object of the present invention is achieved like this:
A kind of integrated capillary biochip, be used for detecting testing sample protein, complementary DNA or RNA, antigen or antibody, medicine or acceptor, polysaccharide or agglutinin, tissue or cell existence whether and what of amount, mainly be made up of bioprobe and carrier element, it is characterized in that: the capillary strip carrier that is provided with the stabilization bioprobe places and constitutes kapillary biochip unit in the capillary cavity; Identical or different kapillary biochip unit is arranged mutually parallel and is bonded into array; Cut above-mentioned kapillary biochip cell array and constitute the integrated capillary biochip along direction vertical or that be not orthogonal to each kapillary biochip unit; Or it is the kapillary biochip cell array after the above-mentioned cutting is mutually bonding again and constitute the integrated capillary biochip.
According to the present invention, its feature also is: by biologic array on the section of the integrated capillary biochip that forms with a kind of kapillary biochip cell array cutting is identical; On its double-edged section, the bioprobe on the pairing corresponding site is identical.
According to the present invention, its feature also is: the identical bioprobe on the described capillary strip carrier is continuous distribution respectively, and different bioprobes is discontinuously arranged; Described capillary strip carrier be solidify again after activating in advance comprise coating, dip-dye, the identical or different bioprobe of growing; The various capillary strip carriers that are preset with bioprobe in capillary cavity are continuously, linking or discontinuous.Described capillary strip carrier is any in capillary fiber, optical fiber, glass fiber, tinsel, the capillary strip film.Described kapillary is based on any in the kapillary of plastics, synthetic material, quartz, glass, metal preparation.
According to the present invention, its feature also is: be filled with filling material in described capillary strip carrier that contains bioprobe and the space between capillary tube inner wall.Described filling material comprises any in plastics, synthetic material, quartz, glass, metal material and the bonding agent thereof.
According to the present invention, its feature also is: described stabilization bioprobe comprises that dna probe, cDNA probe, peptide nucleic acid probe, mRNA, antigen, antibody comprise the monoclonal of different loci or any or arbitrary combination in polyclonal antibody and AIA, medicine or acceptor, polysaccharide or the various biomolecules such as agglutinin, cell or tissue.
According to the present invention, its feature also is: described stabilization bioprobe comprises microballoon and the particulate that is preset with above-mentioned bioprobe, and first-selected is polystyrene microsphere and magnetic particle.Described pre-bag is to be combined on the capillary strip carrier by the polystyrene microsphere of bioprobe or magnetic particle.
According to the present invention, its feature also is: described stabilization bioprobe is any or its composition that contains in sucrose, trehalose, polysaccharide, antiseptic, surfactant, amino acid and the derivant thereof.
According to the present invention, a kind of method for making of integrated capillary biochip is characterized in that:
1) handles activation capillary strip carrier, use 1-ethyl-3-(3-dimethylamino) carbon two Asias again after promptly cleaning the capillary strip carrier
Amine hydrochlorate (EDC) and N-hydroxy thiosuccinimide activators such as (NHS) become capillary strip carrier surface and contain
Reactive group or other reactive groups of active ester are arranged, be beneficial to covalently or non-covalently in conjunction with bioprobe.
2) preparation stabilization bioprobe solution promptly adds sucrose, trehalose, polysaccharide, antiseptic, table in bioprobe solution
Any or its composition in surface-active agent, amino acid and the derivant thereof, the weight ratio of itself and bioprobe is 10-30: 1.Wherein gene probe adopts the synthetic acquisition of dna synthesizer; Peptide nucleic acid probe adopts the synthetic acquisition of Peptide synthesizer; Antigen or antibody probe prepare by reorganization, purifying and monoclonal or polyclonal antibody.3) in each bioprobe solution, put into the capillary strip carrier that has activated, it all is immersed in the bioprobe solution, after treating that curing comprises that coating, dip-dye, growth bioprobe are finished, taking-up is sealed after cleaning, drying, again through being ready for use on next step assembling after cleaning, the drying.4) get the carrier that each root has been preset bioprobe, be placed in the suitable capillary cavity, so repeatedly, prepare identical or different kapillary biochip unit.5) get the kapillary biochip unit that step 4) obtains, be packed in the space between capillary strip carrier and capillary tube inner wall with filling material.6) the identical or different kapillary biochip unit that step 5) is obtained is arranged mutually parallel into array; 7) along vertical or be not orthogonal to the direction cutting step 6 of each kapillary biochip unit) array that obtains and be prepared into the integrated capillary biochip; Or it is the kapillary biochip array after the above-mentioned cutting is mutually bonding again and be prepared into the integrated capillary biochip.According to the present invention, a kind of method for making of integrated capillary biochip, its feature also is;
1) handle all identical capillary strip carriers of activation can be in same container disposable finishing;
2) solidify comprise in conjunction with, all identical bioprobes that are coated with, grow can be in same container disposable finishing;
3) curing comprises that combination, coating, growth bioprobe inequality separately can soak identical capillary strip carrier respectively
Not in the corresponding bioprobe solution in different vessels;
4) the identical bioprobe that cuts on the body of capillary strip is continuous distribution respectively, and different bioprobes is discontinuously arranged;
5) bioprobe on the capillary strip carrier can be to wrap in advance by the polystyrene microsphere of bioprobe and magnetic particle;
6) wrapping by the polystyrene microsphere of bioprobe and magnetic particle in advance is to be combined on the capillary strip carrier;
7) carrier of various default bioprobes all places and constitutes plurality of specifications and polytype kapillary biology in the capillary cavity
Chip unit;
8) carrier of various default bioprobes is continuous, linking or discontinuous in capillary cavity;
9) leave certain space between the carrier of various default bioprobes and capillary cavity inwall;
10) be filled with filling material in the carrier of default bioprobe and the space between the capillary cavity inwall;
11) various kapillary biochips unit can be arranged into plurality of specifications and polytype array according to actual needs;
12) can will be bonded into plurality of specifications and polytype integrated more mutually after above-mentioned array cutting or the cutting according to actual needs
The kapillary biochip;
13) biologic array is identical on the section of same an array of cutting, can a plurality of identical arrays of disposable preparation;
14) on the double-edged section of the same an array that cuts, the bioprobe on the pairing corresponding site is identical;
15) really between various bioprobes in the various arrays realize zero crosstalking, zero pollute.
According to the present invention, a kind of integrated capillary bio-chip test method is characterized in that: optical test, chemogenic deposit, the electronics based on chemical reaction detected, mechanical sense is answered, magnetic induction and Electrochemical Detection; Can carry out the front-back two-sided detection of same integrated capillary biochip; And front-back two-sided testing result is identical; Its image based on optical test is circular or point-like.
According to the present invention, a kind of purposes of integrated capillary biochip is applicable to gene sequencing, medical diagnosis on disease, legal medical expert's evaluation, drug screening, gene expression profile and proteomics research and other life sciences.Beneficial effect
Ingenious design of the present invention is that the capillary strip carrier that is preset with the stabilization bioprobe places formation kapillary biochip unit in the capillary cavity; Identical or different kapillary biochip unit is arranged mutually parallel and is bonded into array; Cut above-mentioned kapillary biochip cell array and constitute the integrated capillary biochip along direction vertical or that be not orthogonal to each kapillary biochip unit; Or it is the kapillary biochip cell array after the above-mentioned cutting is mutually bonding again and constitute the integrated capillary biochip.Advantage and the significant effect more outstanding have been brought therefrom than existing biochip technology.Its advantage and effect are:
1), makes and really between various bioprobes realize zero crosstalking, zero pollute because the capillary strip carrier that will be preset with bioprobe is arranged in the capillary cavity, and the bioprobe on the adjacent capillary cavity internal hair fine strip shape carrier is isolated by adjacent kapillary fully.
2) be located on the capillary strip carrier in various bioprobe stabilizations, and place in the capillary cavity, add and really to realize zero crosstalking, zero pollute, so the present invention has higher stability and accuracy.
3) because on the double-edged section, the bioprobe on the pairing corresponding site is identical, therefore can carry out the front-back two-sided detection of same integrated capillary biochip, and front-back two-sided testing result is identical.Make testing result have comparability, can obviously improve accuracy rate and result's reliability.
4) be bonded into array owing to identical or different kapillary biochip unit is arranged mutually parallel, not only can make in enormous quantities, and respectively be made into all size and various types of kapillary biochip cell array.
5) since the present invention cut above-mentioned kapillary biochip cell array along direction vertical or that be not orthogonal to each kapillary biochip unit to form or the kapillary biochip cell array after the above-mentioned cutting is bonded mutually again, so have high flux, many specifications and eurypalynous characteristics.
6) owing to biologic array on the section of the same an array that cuts is identical, therefore can a large amount of identical arrays of disposable mass preparation, and prepared array has good comparability and practicality, and in like manner prepared the present invention also has good comparability and practicality.
7) since handle all identical capillary strip carriers of activation can be in same container disposable finish and solidify comprise in conjunction with, all identical bioprobes that are coated with, grow can be in same container disposable finishing, so be more suitable for extensive batch making,
And have less difference between batch, therefore have good comparability and practicality.
8) preparation technology of the present invention is easy, is easy to large-scale mass production, has the efficient height, cost is low and advantage such as practical.
9) detection method applicability of the present invention is strong, optical test, chemogenic deposit, the electronics that can be based on chemical reaction detected, mechanical sense is answered, magnetic induction and Electrochemical Detection, and can carry out the front-back two-sided detection of same integrated capillary biochip, resulting front-back two-sided testing result is identical.
What 10) the invention provides a kind of real no crosstalk phenomenon comprises that to gene, protein antigen-antibody, medicine or acceptor, tissue or cell and polysaccharide or agglutinin carry out the good device of parallel detection, identification, evaluation, can be widely used in gene sequencing, medical diagnosis on disease, legal medical expert's evaluation, drug screening, gene expression profile and proteomics research and other life science fields.
Below in conjunction with the drawings and specific embodiments integrated capillary biochip of the present invention and preparation method thereof is described in further detail.
Description of drawings
Fig. 1 is a vertical view of the present invention, and wherein 1 is that kapillary, 2 is that capillary strip carrier, 3 is that bioprobe, 4 is the capillary cavity inner stuffing;
Fig. 2 is an activated carrier method synoptic diagram, and wherein 2 is that capillary strip carrier, 5 is that container, 6 is activator;
Fig. 3 activated carrier is in conjunction with bioprobe method synoptic diagram, and wherein 2a is that container, 7 is bioprobe solution for the capillary strip carrier, 5 that has activated;
Fig. 4 is preparation kapillary biochip cell schematics, and wherein 1 is that kapillary, 2b are for being solidified with the capillary strip carrier of bioprobe;
Fig. 5 is a kapillary biochip cell schematics, and wherein 1 is that kapillary, 2b are that bioprobe, 4 is the capillary cavity inner stuffing for the capillary strip carrier, 3 that has been solidified with bioprobe;
Fig. 6 is the kapillary biochip cell array synoptic diagram of arranging, and wherein 8 is that kapillary biochip unit, 9 is kapillary biochip cell array;
Fig. 7 is a kapillary biochip cell array synoptic diagram, and wherein 9 are the kapillary biochip cell array of kapillary biochip cell array, 10 for having arranged;
Fig. 8 is cutting kapillary biochip cell array synoptic diagram, and wherein 10 are the kapillary biochip array of well cutting for the kapillary biochip cell array of having arranged, 11:
Fig. 9 detects synoptic diagram of the present invention, and wherein 12 is that ccd video camera, 13 is that light filter, 14 is that integrated capillary biochip, 15 is that LASER Light Source, 16 is that computing machine, 17 is data line;
Figure 10 is a detected image synoptic diagram of the present invention
Embodiment
From Fig. 1 to embodiment of the present invention shown in Figure 8 as can be seen, a kind of integrated capillary biochip, mainly be made up of bioprobe and carrier element, it is characterized in that: the capillary strip carrier 2 that is provided with stabilization bioprobe 3 places and constitutes kapillary biochip unit 8 in kapillary 1 chamber; Identical or different kapillary biochip unit 8 is arranged mutually parallel and is bonded into kapillary biochip cell array 10; Cut above-mentioned kapillary biochip cell array 10 and constitute the integrated capillary biochip along direction vertical or that be not orthogonal to each kapillary biochip unit; Or it is the kapillary biochip cell array 11 after the above-mentioned cutting is mutually bonding again and constitute the integrated capillary biochip.
From Fig. 1 or shown in Figure 8, biologic array is identical on the section of the integrated capillary biochip that is formed by same kapillary biochip cell array 10 cuttings; On its double-edged section, the bioprobe 3 on the pairing corresponding site is identical.
From Fig. 2 to embodiment of the present invention shown in Figure 8 as can be seen, it is as follows to prepare method of the present invention:
1) in container 5, handles activation capillary strip carrier 2, use 1-ethyl-3-(3-dimethylamino) carbodiimide hydrochloride (EDC) and N-hydroxy thiosuccinimide activators 6 such as (NHS) that capillary strip carrier 2 surfaces are become reactive group or other reactive groups that contains active ester again after promptly cleaning capillary strip carrier 2, the capillary strip carrier 2a that becomes activation is beneficial to covalently or non-covalently in conjunction with bioprobe.
2) preparation stabilization bioprobe solution 7 in container 5, promptly add any or its composition in sucrose, trehalose, polysaccharide, antiseptic, surfactant, amino acid and the derivant thereof in bioprobe solution 7, the weight ratio of itself and bioprobe is 10-30: 1.Wherein gene probe adopts the synthetic acquisition of dna synthesizer; Peptide nucleic acid probe adopts the synthetic acquisition of Peptide synthesizer; Antigen or antibody probe prepare by reorganization, purifying and monoclonal or polyclonal antibody.
3) in each bioprobe solution 7, put into the capillary strip carrier 2a that has activated, it all is immersed in the bioprobe solution 7, after treating that curing comprises that coating, dip-dye, growth bioprobe are finished, taking-up is sealed after cleaning, drying, again through being ready for use on next step assembling after cleaning, the drying.
4) get the capillary strip carrier 2b that each root has been preset bioprobe, be placed in suitable kapillary 1 chamber, so repeatedly, prepare identical or different kapillary biochip unit 8.
5) get the kapillary biochip unit 8 that step 4) obtains, be packed in the space between capillary strip carrier and kapillary 1 inwall with filling material.
6) the identical or different kapillary biochip unit 8 that step 5) is obtained is arranged mutually parallel into kapillary biochip cell array 10;
7) along vertical or be not orthogonal to the direction cutting step 6 of each kapillary biochip unit 8) the kapillary biochip cell array 10 that obtains and be prepared into the integrated capillary biochip; Or it is the kapillary biochip array 11 after the above-mentioned cutting is mutually bonding again and be prepared into the integrated capillary biochip.
Find out that from Fig. 9 and detection embodiments of the present invention shown in Figure 10 this integrated capillary bio-chip test method is: based on the optical test of chemical reaction.Under the irradiation of LASER Light Source 15, the image that is produced is by ccd video camera 12 picked-ups with this integrated capillary biochip 14 of finishing chemical reaction, and wherein the image of optical test is circular or point-like.By the biochip image analysis software on the computing machine 16, can obtain the optical test result of biochip then.And can carry out the front-back two-sided detection of same integrated capillary biochip 14, and front-back two-sided testing result is identical.
During use, the sample to be checked of getting 1-2m1 adds on the side section of the present invention or with the present invention invades in the sample to be checked.The present invention is owing to contain bioprobe 3 simultaneously on its double-edged section, after adding sample to be checked, 37 ℃ question response 2-60 minute, specific biochemistry or immunity or complementary interaction take place in the thing component to be checked in bioprobe in the bioprobe 3 and the sample to be checked, get cleansing solution washing the present invention then, give a baby a bath on the third day after its birth altogether time, wash away the material that does not react, add label reagent then and comprise metal marker, enzyme labeling, fluorescence labeling, chemiluminescent labeling, the electrochemiluminescence mark, dye marker and radioactive label reagent, 37 ℃ were reacted 2-60 minute, label and the reaction of above-mentioned specific complex, get cleansing solution washing the present invention then, give a baby a bath on the third day after its birth altogether time, wash away the label that does not react, add developer subsequently, question response passes through the chromogenic reaction sentence read result at last, or washs back sentence read result on laser scanner or confocal laser scanning microscope, CLSM or CCD chip scanner or other equipment with cleansing solution, colour developing or luminous positive, colourless or not luminous negative.If corresponding thing component to be checked is arranged in the sample to be checked, colour developing or luminescence-producing reaction take place with regard to energy specificity binding label reagent, the result is positive; If there is not corresponding thing component to be checked in the sample to be checked, just can not specificity binding label reagent, label reagent is washed to be removed, and colour developing or luminescence-producing reaction just can not take place, and the result is negative.
Genetic chip among the present invention is a kind of special shape of the present invention, its key distinction is: label reagent is the label of DNA in the testing sample, can be labels such as metal marker, enzyme labeling, fluorescence labeling, chemiluminescent labeling, electrochemiluminescence mark, dye marker and radioactive label.After adding sample to be checked, dna marker thing in the sample to be checked and the bioprobe in the present invention comprise that complementation takes place for dna probe, cDNA probe, peptide nucleic acid probe and mRNA, be used for showing the sample to be checked of being caught by bioprobe the complementary DNA label existence whether.Its concrete operations are: after adding sample DNA label to be checked, 37 ℃ were reacted 30-60 minute, bioprobe comprises dna probe in sample DNA label to be checked and the present invention, the cDNA probe, complementation takes place in peptide nucleic acid probe and mRNA, get cleansing solution washing the present invention then, give a baby a bath on the third day after its birth altogether time, wash away the material that does not react, or add the developer question response subsequently, colour developing or luminescence-producing reaction sentence read result by the dna marker thing, or with direct sentence read result on laser scanner or confocal laser scanning microscope, CLSM or CCD chip scanner or other equipment after the cleansing solution washing, colour developing or luminous positive, colourless or not luminous negative.If the existence of complementary DNA or RNA is arranged in the sample to be checked, remove with regard to can specificity being combined in can not be washed in the present invention, colour developing or luminescence-producing reaction take place then, the result is positive; If the existence of no complementary DNA or RNA in the sample to be checked just can not specificity be combined in to be washed in the present invention and removes, colour developing or luminescence-producing reaction just can not take place, the result is negative.
Claims (10)
1, a kind of integrated capillary biochip mainly is made up of bioprobe and carrier element, it is characterized in that: the capillary strip carrier that is provided with the stabilization bioprobe places and constitutes kapillary biochip unit in the capillary cavity; Identical or different kapillary biochip unit is arranged mutually parallel into kapillary biochip cell array; Cut above-mentioned kapillary biochip cell array and constitute the integrated capillary biochip along direction vertical or that be not orthogonal to each kapillary biochip unit; Or it is the kapillary biochip cell array after the above-mentioned cutting is mutually bonding again and constitute the integrated capillary biochip.
2, integrated capillary biochip according to claim 1 is characterized in that: biologic array is identical on the section of the integrated capillary biochip that is formed by same kapillary biochip cell array cutting; On its double-edged section, the bioprobe on the pairing corresponding site is identical.
3, integrated capillary biochip according to claim 1 and 2 is characterized in that: the identical bioprobe on the described capillary strip carrier is continuous distribution respectively, and different bioprobes is discontinuously arranged; Described capillary strip carrier be solidify again after activating in advance comprise coating, dip-dye, the identical or different bioprobe of growing; The various capillary strip carriers that are preset with bioprobe in capillary cavity are continuously, linking or discontinuous.Described capillary strip carrier is any in capillary fiber, optical fiber, glass fiber, tinsel, the capillary strip film.Described kapillary is based on any in the kapillary of plastics, synthetic material, quartz, glass, metal preparation.
4, integrated capillary biochip according to claim 1 and 2, its feature also is: be filled with filling material in described capillary strip carrier that contains bioprobe and the space between capillary tube inner wall.Described filling material comprises any in plastics, synthetic material, quartz, glass, metal material and the bonding agent thereof.
5, integrated capillary biochip according to claim 1 and 2 is characterized in that: described stabilization bioprobe comprises that dna probe, cDNA probe, peptide nucleic acid probe, mRNA, antigen, antibody comprise any or the arbitrary combination in the monoclonal of different loci or polyclonal antibody and AIA, medicine or acceptor, polysaccharide or agglutinin, the cell or tissue.
6, integrated capillary biochip according to claim 4 is characterized in that: described stabilization bioprobe comprises microballoon and the particulate that is preset with above-mentioned bioprobe, and first-selected is polystyrene microsphere and magnetic particle.Described pre-bag is to be combined on the capillary strip carrier by the polystyrene microsphere of bioprobe or magnetic particle.
7, the method for making of integrated capillary biochip is characterized in that:
1) handles activation capillary strip carrier, use 1-ethyl-3-(3-dimethylamino) carbodiimide hydrochloride and N-hydroxy thiosuccinimide that capillary strip carrier surface is become the reactive group that contains active ester again after promptly cleaning the capillary strip carrier, be beneficial to covalently or non-covalently in conjunction with bioprobe.
2) preparation stabilization bioprobe solution promptly adds any or its composition in sucrose, trehalose, polysaccharide, antiseptic, surfactant, amino acid and the derivant thereof in bioprobe solution, the weight ratio of itself and bioprobe is 10-30: 1.Wherein gene probe adopts the synthetic acquisition of dna synthesizer, and antigen or antibody probe prepare by abundance, purifying and monoclonal or polyclonal antibody.
3) in each bioprobe solution, put into the capillary strip carrier that has activated, it all is immersed in the bioprobe solution, after treating that curing comprises that coating, dip-dye, growth bioprobe are finished, taking-up is sealed after cleaning, drying, again through being ready for use on next step assembling after cleaning, the drying.
4) get the carrier that each root has been preset bioprobe, be placed in the suitable capillary cavity, so repeatedly, prepare identical or different kapillary biochip unit.
5) get the kapillary biochip unit that step 4) obtains, be packed in the space between capillary strip carrier and capillary tube inner wall with filling material.
6) the identical or different kapillary biochip unit that step 5) is obtained is arranged mutually parallel into array;
7) along vertical or be not orthogonal to the direction cutting step 6 of each kapillary biochip unit) array that obtains and be prepared into the integrated capillary biochip; Or it is the kapillary biochip array after the above-mentioned cutting is mutually bonding again and be prepared into the integrated capillary biochip.
8, the method for making of integrated capillary biochip according to claim 6 is characterized in that:
1) handle all identical capillary strip carriers of activation can be in same container disposable finishing;
2) solidify comprise in conjunction with, all identical bioprobes that are coated with, grow can be in same container disposable finishing;
3) solidify and to comprise in conjunction with, coating, growth bioprobe inequality separately and identical capillary strip carrier can be immersed in respectively in the corresponding bioprobe solution in the different vessels;
4) the identical bioprobe on the capillary strip carrier is continuous distribution respectively, and different bioprobes is discontinuously arranged.
5) bioprobe on the capillary strip carrier can be to wrap in advance by the polystyrene microsphere of bioprobe and magnetic particle;
6) wrapping by the polystyrene microsphere of bioprobe and magnetic particle in advance is to be combined on the capillary strip carrier;
7) carrier of various default bioprobes all places and constitutes plurality of specifications and polytype kapillary biochip unit in the capillary cavity;
8) carrier of various default bioprobes is continuous, linking or discontinuous in capillary cavity;
9) leave certain space between the carrier of various default bioprobes and capillary cavity inwall;
10) various kapillary biochips unit can be arranged into plurality of specifications and polytype array according to actual needs;
11) can will be bonded into plurality of specifications and polytype integrated capillary biochip more mutually after above-mentioned array cutting or the cutting according to actual needs;
12) biologic array is identical on the section of same an array of cutting, can a plurality of identical arrays of disposable preparation;
13) on the double-edged section of the same an array that cuts, the bioprobe on the pairing corresponding site is identical;
14) really between various bioprobes in the various arrays realize zero crosstalking, zero pollute.
9, according to claim 1 or 7 described integrated capillary bio-chip test method, it is characterized in that: optical test, chemogenic deposit, the electronics based on chemical reaction detected, mechanical sense is answered, magnetic induction and Electrochemical Detection; Can carry out the front-back two-sided detection of same integrated capillary biochip, and front-back two-sided testing result is identical; Its image based on optical test is circular or point-like.
10, according to the purposes of claim 1 or 7 described integrated capillary biochips, it is characterized in that: this integrated capillary biochip is applicable to gene sequencing, medical diagnosis on disease, legal medical expert's evaluation, drug screening, gene expression profile and proteomics research and other life sciences.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 03141391 CN1458525A (en) | 2003-06-08 | 2003-06-08 | Integrated capillary biological chip and its producing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN 03141391 CN1458525A (en) | 2003-06-08 | 2003-06-08 | Integrated capillary biological chip and its producing method |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1312293C (en) * | 2004-11-08 | 2007-04-25 | 博奥生物有限公司 | High flux biochip and application thereof |
| CN108344697A (en) * | 2018-05-14 | 2018-07-31 | 苏州大学 | A kind of special loading shelf combination of the external sample of optoacoustic tomoscan molecular imaging system |
| CN110420671A (en) * | 2019-06-26 | 2019-11-08 | 南京科瑞芯生物科技有限公司 | A kind of agglutinin micro-array chip and preparation method thereof |
| CN110632287A (en) * | 2019-08-15 | 2019-12-31 | 成都工业学院 | Capillary tube containing probe array and application and preparation method thereof |
| CN114181804A (en) * | 2022-02-15 | 2022-03-15 | 深圳市研元生物科技有限公司 | Gene chip and preparation and use method thereof |
-
2003
- 2003-06-08 CN CN 03141391 patent/CN1458525A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1312293C (en) * | 2004-11-08 | 2007-04-25 | 博奥生物有限公司 | High flux biochip and application thereof |
| CN108344697A (en) * | 2018-05-14 | 2018-07-31 | 苏州大学 | A kind of special loading shelf combination of the external sample of optoacoustic tomoscan molecular imaging system |
| CN108344697B (en) * | 2018-05-14 | 2024-02-09 | 苏州大学 | Special sample loading frame combination for in-vitro samples of photoacoustic tomography molecular imaging system |
| CN110420671A (en) * | 2019-06-26 | 2019-11-08 | 南京科瑞芯生物科技有限公司 | A kind of agglutinin micro-array chip and preparation method thereof |
| CN110632287A (en) * | 2019-08-15 | 2019-12-31 | 成都工业学院 | Capillary tube containing probe array and application and preparation method thereof |
| CN114181804A (en) * | 2022-02-15 | 2022-03-15 | 深圳市研元生物科技有限公司 | Gene chip and preparation and use method thereof |
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