CN1242071C - Drum type technique for preparing biochip in micro array mode - Google Patents

Abstract

The present invention provides a drum type fabrication technology of a micro array biochip. According to probes which are distributed on a biochip to be prepared, materials which are modified with probe molecules and have built-in pore structures are selected and are fabricated into a straight line shape. According to the distribution condition of the probes on the biochip to be prepared, the materials which are modified with the probe molecules and have the built-in pore structures are parallel paved on the other flexible solid base sheet. After the surface of the base sheet is coated with a bonding agent, the flexible solid base sheet is reeled in the paving direction of the straight line-shaped materials which are modified with the probe molecules, which enables the straight line-shaped materials which are modified with the probe molecules and have the built-in pore structures to be clamped at different positions of the reeled base sheet along with the reeling until the base sheet is reeled into a reel drum shape. In a perpendicular direction of the straight line-shaped material which are modified with the probe molecules and have the built-in pore structures, the base sheet of the reel drum shape is mechanically cut, and an obtained sheet is a micro array biochip. The present invention has the advantages of standardization and mass production and low preparing cost.

Description

The roll type manufacturing process of micro-array biochip

Technical field

The present invention relates to a kind of preparation method of biochip, relate in particular to a kind of roll type manufacturing process of micro-array biochip.

Background technology

Biochip mainly refers to the microarray that the biologically active substance (comprising nucleic acid, protein, cell and small tissue etc.) assembled constitutes in the solid chip, with realize to compound (comprising medicine), protein, nucleic acid, cell and other biological components accurately, fast, the screening or the detection of large information capacity.The principal feature of biochip comprises: (1) can assemble a large amount of (10 in a less solid chip ²~ 10 ⁶Kind) biologically active substance, that obtains contains much information, the efficient height, and the biological reagent of consumption is few, can save a large amount of reagent expenses, and cost is low; (2) adopted plane and three-dimensional Micrometer-Nanometer Processing Technology, can reduce the preparation cost of single chip, realized producing in enormous quantities by improving integrated level; (3) can the pre-treatment of biological sample be extracted, amplification, reaction, and the information detection is mutually integrated, is prepared into miniature, full-automatic, pollution-free, as the to can be used for the microsample detection integrated intelligent biology chip of height.

Biochip has extremely important meaning on biological detection, medical test and medical diagnosis on disease, drug screening and gene sequencing.Traditional biological detects the method adopted and comprises a series of numerous and diverse steps, especially extensive biological compound detect and screening aspect time-consuming, effort, cost height, can not satisfy the demand.Traditional method is being carried out in the improved process, is that the biochip technology of representative arises at the historic moment with the gene chip.The strong means that provide are obtained and analyzed to the maturation of this technology and use and will bring a revolution for the exploitation and the life science association areas such as evaluation, food and environment of new drug for bioinformation.

At present, biochip has two kinds of main preparation methods: a few point sample preparations and original position are synthetic, but these two kinds of methods all are not suitable for large-scale production.

Technology contents

The invention provides a kind of roll type manufacturing process of micro-array biochip, have preparation cost low, the operation simple and feasible, the advantage of can stdn and producing quantitatively, compound molecule density is even on the lattice point of the array chip that is made by the present invention, makes the accuracy of chemical array chip analysis higher.

The present invention adopts following technical scheme:

The first step: according to the probe that is distributed on the biochip to be prepared, choose the material that includes pore structure of having modified these probe molecules, be made into rectilinear form,

Second step: according to the probe distribution situation on the biochip to be prepared, pave on another soft solid substrate the material that includes pore structure that is modified with probe molecule of above-mentioned rectilinear form is parallel,

The 3rd step: behind substrate surface brushing binding agent, along with the direction scrolling soft solid substrate that paves of the rectilinear form material that is modified with probe molecule, make the rectilinear form material that includes pore structure that is modified with probe molecule be sandwiched between the different sites of volume mera with this scrolling, be rolled into bobbin shape until substrate

The 4th the step: along with the perpendicular direction of rectilinear form material that includes pore structure that is modified with probe molecule, the substrate of cut mechanically bobbin shape, resulting thin slice is micro-array biochip.

Compared with prior art, the present invention has following advantage:

1) because block is carried out cut mechanically, can obtain many identical porous membranes simultaneously, then obtain many identical porous membranes that are fitted in the glass slide surface, according to the needs of classes of compounds on the micro-array chip of chemical compound to be prepared and distribution thereof, can prepare many identical or different compound chips.Thereby, can stdn and produce quantitatively, preparation cost is low;

2) because the straight line probe of arranging in the plane is easy to operate, be not easy to make the mistake, can be isodensity, also can be that non-isodensity is arranged, do not need expensive equipment, the making ability of chemical array chip be greatly enhanced simultaneously;

3) owing to the straight line probe of arranging in the plane, what do not meet the requirements can reject, and resulting compound molecule lattice point marshalling is convenient to scanning analysis.

4) because the present invention can utilize simple winding apparatus, as: become bobbin shape by will the pave soft solid web substrate of probe molecule of rotating drum, this method cost is lower, operation is simpler and easy, and stdn and mass production degree are higher.

Specific embodiments

The roll type manufacturing process of 1 one kinds of micro-array biochips of embodiment: the first step: according to the probe that is distributed on the biochip to be prepared, choose the material that includes pore structure of having modified these probe molecules, be made into rectilinear form, this rectilinear form material that includes pore structure that is modified with probe molecule can adopt with not with the inorganic materials of probe molecule generation chemical reaction, organic materials or its matrix material, as: glass, quartzy, pottery, crystal, metal etc., rubber, plastics, gel, nylon, biomacromolecule, macromolecular materials such as poly-electric polymer, make the straight line kapillary, with its internal surface activation, again with oligonucleotide, polypeptide, probe molecules such as biomacromolecule such as polysaccharide and peptide nucleic acid(PNA) and mixture thereof are incorporated into straight line inwall capillaceous, in addition, present embodiment can adopt following method to obtain to be modified with the rectilinear form material that includes pore structure of probe molecule: 1. select porous material for use and modify probe molecule in the hole of porous material, obtain being modified with the material that includes pore structure of probe molecule; 2. select Web materials for use and in the mesh of Web materials, modify probe molecule, obtain being modified with the material that includes pore structure of probe molecule; 3. select glass capillary for use, in the inwall of glass capillary plating reflecting layer, modify probe molecule in glass capillary, what obtain being modified with probe molecule includes pore structure rectilinear form material.For example, obtain containing the surface of active group amino after glass material reacts by the 3-TSL 8330, and the active amino on surface can further obtain the surface of active group aldehyde radical with the glutaraldehyde reaction.And the compound that the surface of containing active group amino can be modified with phosphate radical (as oligonucleotide etc.) reacting makes compound molecule be fixed at glass surface; And contain the active group aldehyde radical the surface can reacting makes compound molecule be fixed at glass surface with containing amino compound (as amido modified oligonucleotide, protein etc.).

Second step: according to the probe distribution situation on the biochip to be prepared, pave on another soft solid substrate the material that includes pore structure that is modified with probe molecule of above-mentioned rectilinear form is parallel, substrate material is optional to be used in and bent by the external force effect and do not produce the organic materials of fracture, as: macromolecular materials such as rubber, plastics, gel, nylon, biomacromolecule, poly-electric polymer.

The 3rd step: behind substrate surface brushing binding agent, along with the direction scrolling substrate that paves of the rectilinear form material that includes pore structure that is modified with probe molecule, make the rectilinear form material that includes pore structure that is modified with probe molecule be sandwiched between the different sites of volume mera with this scrolling, be rolled into bobbin shape until substrate

The 4th the step: along with the perpendicular direction of rectilinear form material that includes pore structure that is modified with probe molecule, the substrate of cut mechanically bobbin shape, resulting thin slice is micro-array biochip.

The roll type manufacturing process of 2 one kinds of micro-array biochips of embodiment:

The preparation method one of A, roll type compound microarray,

A) have at one and arranged some hollow microchannels such as small Glass tubing, metal tube, polymer tube etc. on the certain thickness soft solid substrate, be filled with netted or porous material in the microchannel; Compound molecule combines with some hollow microchannels by chemical bonding, mode such as UV-crosslinked; Then these plane lattices of being made up of some open tubes are obtained the little battle array of folded formula compound by being superimposed upon under the adhesive bond effect; Then these linear lattices of being made up of some open tubes are bonded to roll type compound microarray by rolling volume;

B) use rapid section machinery, block materials along being cut into parallel thin slice with the vertical direction of microtubule with certain angle, is promptly made required film chip;

The preparation method two of B, roll type compound microarray

A) one have be arranged in parallel on the certain thickness soft solid substrate some by being a kind of during the rectilinear tubes (rod, rope) that comprises the compound molecule of the chemical action bonding between hydroxyl, amino, aldehyde radical, carboxyl etc. waits by physical fixation such as UV-crosslinked, absorption or active group or making up; Then these compound plane lattices of being made up of some linear arrays are prepared roll type compound microarray by rolling the volume bonding;

B) use rapid section machinery, block materials along being cut into parallel thin slice with the vertical direction of linear lattice, is promptly made required film chip;

The preparation method three of C, roll type compound microarray

A) have on the certain thickness soft solid substrate at one, the inorganic materials modification of surfaces that adopts macromolecular material or melt is at the different bioactive molecule of form connection of these surperficial different zones with linear array; Then these compound plane lattices of being made up of some linear arrays are prepared roll type compound microarray by rolling the volume bonding;

B) use rapid section machinery, block materials along being cut into parallel thin slice with the vertical direction of linear lattice, is promptly made required film chip;

(2) fixed form of compound

The fixed form one of A, compound:

By modes such as UV-crosslinked, physical adsorptions, specific compound or biomolecules are fixed in the netted and porous material that microtubule fills; Or at specific compound or the biological substance of some regional directly regulation on substrate material.

The fixed form two of B, compound:

By the mode of combinatorial chemistry, directly synthetic specific compound or biological substance in porous in microtubule or the Web materials; Or at directly synthetic specific compound or the biological substance in some zone on substrate material.

Compound embedding or physical adsorption are prepared into roll type compound microarray in microtubule and linear lattice zone and with it, behind the section film forming chip, under the effect of light, electricity, heat or magnetic, make compound molecule and propping material chemical bonding.

The fixed form three of C, compound:

By the active group on the compound molecule is to comprise the mode of the group chemical bonding on hydroxyl, amino, aldehyde radical, carboxyl etc. and the carrier or by mode UV-crosslinked, physical adsorption, directly connect compound molecule, make the zone that connects compound molecule constitute the compound plane lattice in some specific region of soft substrate material surface.

(3) with in the preparation form (1) of above-mentioned plane micro-array chip any, make up with in the fixed form of above-mentioned (2) compound any, the compound microarray that obtains just can be prepared roll type compound microarray by rolling the volume bonding, this roll type compound microarray promptly makes required film chip by the machinery section.

The preparation of embodiment 3 oligonucleotide arrays film chips

Chemically modified aldehyde radical on the different zones of soft solid substrate, combine by firm chemical bond with amido modified oligonucleotide, make the oligonucleotide that directly secured in parallel is different on the soft solid substrate in the different zone of chemically modified constitute oligonucleotide plane microarray, then this block materials is bonded to the roll type oligonucleotide microarray along the direction clove hitch parallel with the oligonucleotide zone, last along thinly slicing with the direction of small oligonucleotide regions perpendicular, constitute oligonucleoside sorrel chip.

Embodiment 4 genomic dnas or the preparation of cDNA library film chip

Different size gene or cDNA fragment are introduced different packed columns, by the chemical bond key and, perhaps make it to be fixed in the packed column by ultraviolet light cross-linking, then packed column being sticked with glue agent is fixed on the soft solid material, be bonded to roll type genomic dna or cDNA library microarray along this block materials along direction clove hitch with small cartridges parallel, last along thinly slicing with the vertical direction of small cylinder, constitute genomic dna or cDNA library film chip.

The preparation of embodiment 5 albuminous membranae chips

Amino in the aldehyde group modified different zone of soft solid substrate and protein constitutes protein plane microarray by chemical bond in conjunction with the different protein of secured in parallel on, the soft solid substrate in chemically modified, then this block materials is bonded to the roll type protein microarray along the direction clove hitch parallel with the protein zone, last along thinly slicing with the direction of small protein regions perpendicular, constitute the albuminous membranae chip.

Claims (1)

1, a kind of roll type manufacturing process of micro-array biochip is characterized in that:

The first step: according to the probe that is distributed on the biochip to be prepared, choose the material that includes pore structure of having modified these probe molecules, be made into rectilinear form,

Second step: according to the probe distribution situation on the biochip to be prepared, pave on another soft solid substrate the material that includes pore structure that is modified with probe molecule of above-mentioned rectilinear form is parallel,

The 3rd step: behind substrate surface brushing binding agent, along with the direction scrolling soft solid substrate that paves of the rectilinear form material that is modified with probe molecule, the rectilinear form material that includes pore structure that is modified with probe molecule is rolled up between the different sites of mera by clip with this scrolling, be rolled into bobbin shape until substrate

The 4th the step: along with the perpendicular direction of rectilinear form material that includes pore structure that is modified with probe molecule, the substrate of cut mechanically bobbin shape, resulting thin slice is micro-array biochip.