CN1314476C - Method and equipment for purifying and recovering biomacromolecules - Google Patents
Method and equipment for purifying and recovering biomacromolecules Download PDFInfo
- Publication number
- CN1314476C CN1314476C CNB2005100242571A CN200510024257A CN1314476C CN 1314476 C CN1314476 C CN 1314476C CN B2005100242571 A CNB2005100242571 A CN B2005100242571A CN 200510024257 A CN200510024257 A CN 200510024257A CN 1314476 C CN1314476 C CN 1314476C
- Authority
- CN
- China
- Prior art keywords
- electrode
- purifying
- electrophoresis
- pipeline
- sample
- 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.)
- Expired - Fee Related
Links
Abstract
The present invention relates to a method and a device for purifying and recovering biomacromolecules. In the method, biomacromolecules to be recovered and purified are led into collecting pits by the selectivity switching of electrophoresis electrodes in an electrophoresis mode, and therefore, the recovery and the purification of the biomacromolecules are accomplished. Voltage is loaded on electrodes on both ends of a main electrophoresis passage so that samples are separated on the main electrophoresis passage. Meanwhile, the positions of streaming biomacromolecule strips are monitored through an optical detecting method, and after the biomacromolecule strips to be recovered and purified arrive at the proper positions, the biomacromolecules to be recovered and purified are led into a nearest branch pipeline through properly switching the electrodes and are finally led into the corresponding collecting pits. The present invention provides a device for once accomplishing the separation and the purification of the biomacromolecules, and the device has the characteristics of simple structure, easy operation and easy realization of integration and automation. The present invention can be applied to purifying and recovering nucleic acid, protein, a nucleic acid-protein compound, etc. in molecular biology.
Description
Technical field
The present invention relates to method and device thereof that a kind of biological macromolecule purifying reclaims.The method and device can be applicable to molecular biological field.
Background technology
Large biological molecules such as DNA, protein, DNA-albumen composition to cut that glue reclaims be a routine operation of biology laboratory, cut the purifying of product, marked product etc., the separation and purification of marking protein, DNA-albumen composition, RNA-albumen composition etc. etc. such as PCR product, enzyme.
At present, the method that the dna molecular purifying reclaims mainly contains two kinds, a kind of is conventional laboratory method: electrophoresis → cut glue → melt glue → phenol chloroform extracting → precipitation with alcohol → dissolving, this method complex operation step, technology and qualification to operating personnel have certain requirement, and include and poison reagent, and have the quantity of sample loss, thereby be unsuitable for the purifying recovery of micro-example.Another kind method is the commercially available reagent cassette method: electrophoresis → cut glue → melt glue → centrifugal is crossed post → wash-out dissolving, this method equally also has certain specification requirement to operating personnel, there is the quantity of sample loss equally, purifying recovery to micro-example is unfavorable, and existing kit price is expensive, be difficult for popularizing [Sheng Xiaoyu translates .DNA and RNA basic experiment technology. Beijing: the .2002.Frederick Ausubel et al.ShortProtocols in Molecular Biology.John Wiley of Science Press; Sons, Inc., Publishers, 2000].
At present, the method for purifying and recycling of protein molecule mainly contains several different methods [Richard J.Simpson.Purifyingproteins for proteomics:a Laboratory Manual.Cold Spring HarborLab (CSHL) Press such as the precipitation method, gel filtration, electrophoresis, ion exchange chromatography, affinity chromatography, 2004], the complicated operation that these methods have, consuming time longer, the organic efficiency that has is low, and causes albuminous degeneration easily, and what have needs cryogenic conditions.Lack a kind of easy and simple to handlely, albumen reclaims purification process fast and effectively.
Summary of the invention
The purpose of this invention is to provide a kind of method of biological macromolecule purifying recovery and the separating and the one step completed device of purifying large biological molecule of Using such method design, simplify the operating procedure that biological macromolecule purifying reclaims, the use of poisonous chemical reagent in avoiding operating, improve the rate of recovery of sample simultaneously, reduce the sample cost recovery, and provide a kind of simple in structure, with low cost, easy and simple to handle, easily be automated, and can be used for the biological macromolecule purifying retracting device of microsystem.
The method that a kind of biological macromolecule purifying provided by the invention reclaims, it is characterized in that this method utilizes the selectivity of iontophoretic electrode to switch, at first large biological molecule is separated by the electrophoresis mode, large biological molecule with purifying to be recycled imports collecting pit then, thereby realizes the recovery purifying of large biological molecule.
Particularly, when implementing the recovery of sample purifying, at first on-load voltage on the electrode that is positioned at the main track two ends makes sample be separated on main track.Monitor the position of swimming large biological molecule band simultaneously by optical detecting method, after the large biological molecule band of purifying to be recycled and other band are separated fully, by optionally switching electrode, the large biological molecule of purifying to be recycled is introduced nearest lateral, and the final collecting pit that imports correspondence, the purifying of finishing sample reclaims.Whole electrophoresis process relates to the selectivity switching of plural electrode and electrode; The switching of electrode is by the electrophoretic band determining positions of the large biological molecule of recovery to be purified.In the electrophoresis pipeline, fill solid-state sieving media, as Ago-Gel; Charge into electrophoresis cushioning liquid in the addition pool of duct end and the collecting pit, as TBE.
The present invention also provides a kind of device of implementing above-mentioned biological macromolecule purifying recovery method, this device comprises the electrophoresis parts that are processed with branch shape pipeline and some electrodes, the monitoring component of the big molecule electrophoretic band of monitoring bio position and control electrode ON-OFF control circuit parts.It is characterized in that: its electrophoresis parts comprise a main separating pipe and one or more lateral, and electrode is all arranged in the Xiao Chi of each duct end; Electrode can contact with buffer solution among the Xiao Chi by inserted mode, also can by sputter or electroplating processing process be fixed in the Xiao Chi bottom directly and the buffer solution among the Xiao Chi contact.Be positioned at the electrode of collecting pit or outside and coat one deck pellicle, perhaps and isolate the material of one deck pellicle between the lateral.
The present invention compares with biological macromolecule purifying recovery method commonly used at present, simplified the operating procedure that biological macromolecule purifying reclaims, avoid the use of poisonous chemical reagent in the operation, improved the rate of recovery of sample simultaneously, reduced the cost that the sample purifying reclaims.In addition, apparatus of the present invention are simple in structure, and easy operating is easy to realize integrated and automation.
Description of drawings
Fig. 1 is apparatus of the present invention schematic diagram.
Fig. 2 is the electrophoresis parts stereochemical structure view of the embodiment of the invention.
Fig. 3 is an electrophoresis parts bottom electrode substrate plane view shown in Figure 2.
Fig. 4 is the electrophoresis parts intermediate conduit shown in Figure 2 and the intermediate layer plan view of cavity.
Fig. 5 is an electrophoresis parts upper strata cover plate plan view shown in Figure 2.
The specific embodiment
Further specify substantive distinguishing features of the present invention and obvious improvement below in conjunction with accompanying drawing.
Fig. 1 is apparatus of the present invention schematic diagram, the utilization of this device has the swimming that the electrophoresis parts 1 of branch shape electrophoresis pipeline and specific configuration electrode drive large biological molecule in the sample and separates, position by electrophoretic band in the monitoring component 2 monitoring electrophoresis parts 1, and feed back to control assembly 3, utilize the switch and the alive size of each electrode on the control circuit control electrophoresis parts 1 in the control assembly 3.
Fig. 2 is the electrophoresis parts stereochemical structure view of the embodiment of the invention.These parts comprise three-decker, and the substrate 4 that is processed with the specific configuration electrode is positioned at bottom, and the intermediate layer is the pipe layers 5 that is processed with branch shape electrophoresis pipeline and cavity, and the superiors are the cover plate 6 that is processed with corresponding aperture.
Fig. 3 is the electrophoresis parts bottom electrode substrate plane view of the embodiment of the invention.The substrate zone line is placed with realizes separation and each electrode of collecting, is followed successively by keeper electrode 9, passive electrode 10 and the main separate mesh electrode 12 of application of sample electrode 7, band shaping electrode 8, recovery sample.The periphery be being connected of drawing of each electrode with control circuit be connected electrode 11.
Fig. 4 has the pipe layers plan view of branch shape electrophoresis pipeline and cavity for the electrophoresis component processing of the embodiment of the invention.According to the function difference, purifying reclaims electrophoresis pipeline 14 and has difformity with reference to electrophoresis pipeline 15.With reference to electrophoresis pipeline 15 is single straight pipeline, and two ends have addition pool 13 and waste liquid pool 17 respectively; Purifying reclaims electrophoresis pipeline 14 except the addition pool 13 and waste liquid pool 17 at main separating pipe two ends, then than reference electrophoresis pipeline 15 more laterals and collecting pit 16, main separating pipe has three laterals and corresponding collecting pit in the present embodiment as shown in the figure.
Fig. 5 is the electrophoresis parts upper strata cover plate plan view of the embodiment of the invention.The cover plate layer comprises the well 18 of being convenient to application of sample and sampling, collection hole 19 and waste liquid hole 20.The position of well 18 should be corresponding to the addition pool in the pipe layers 5 13, and collection hole 19 should reclaim lateral and collecting pit 16 on the electrophoresis pipeline 14 corresponding to purifying, and waste liquid hole 20 should be corresponding to the waste liquid pool on the pipe layers 3 17.
When implementing the recovery of sample purifying, at first in two wells 18 of electrophoresis parts 1, add sample respectively, reclaim the sample liquid that adds recovery to be purified in the well that electrophoresis pipeline 14 communicates with purifying, with add reference sample liquid in the well that reference electrophoresis pipeline 15 communicates, between application of sample electrode 7 and sample strip shaping electrode 8, load the voltage of suitable direction and intensity then, large biological molecule in the sample liquid is enriched in rapidly on the sample strip shaping electrode 8, forms the sample strip that elongated and narrow is easy to differentiate.After finishing the sample strip shaping, switch electrode, between application of sample electrode 7 and main separate mesh electrode 12, load the voltage of suitable direction and intensity, make sample on main track, be separated.Monitor the position of the large biological molecule band of swimming simultaneously by optical detecting method, after the large biological molecule band of purifying to be recycled arrives suitable position, by optionally switching electrode, the large biological molecule of purifying to be recycled is enriched on the nearest keeper electrode 9.The last voltage that loads suitable direction and intensity between the recovery electrode 10 of keeper electrode 9 and respective branches collection conduit terminal imports corresponding collecting pit with the large biological molecule band to be recycled that is enriched on the keeper electrode 9, and the purifying of finishing sample reclaims.After this process is finished the separation of large biological molecule, directly large biological molecule to be recycled is imported in the buffer solution, saved and cut glue in the normal experiment method, melted glue, the step of the purifying that removes photoresist then.Collected large biological molecule sample can be directly used in follow-up molecular biology operation, as order-checking, clone, mark, amplification, restriction enzyme digestion and in-vitro transcription translation etc.Obvious retracting device provided by the invention will separate and purifying is once finished, and be better than existing method and apparatus.
Claims (8)
1. the method that biological macromolecule purifying reclaims it is characterized in that described method is to utilize the selectivity of iontophoretic electrode to switch by the large biological molecule importing collecting pit of electrophoresis mode with purifying to be recycled, thereby the purifying of realizing large biological molecule reclaims.Specifically be on-load voltage on the electrode that is positioned at the main track two ends at first, make sample on main track, be separated; Monitor the position of swimming large biological molecule band simultaneously by optical detecting method, after the large biological molecule band of purifying to be recycled and other band are separated fully, by optionally switching electrode, the large biological molecule of purifying to be recycled is introduced nearest lateral, and the final collecting pit that imports correspondence, the purifying of finishing sample reclaims.
2. according to the method for the described biological macromolecule purifying recovery of claim 1, it is characterized in that: whole electrophoresis process relates to the selectivity switching of plural electrode and electrode; The switching of electrode is by the electrophoretic band determining positions of the large biological molecule of recovery to be purified.
3. implement biological macromolecule purifying retracting device as claimed in claim 1, it is characterized in that: this device is by being processed with the branch shape pipeline and the electrophoresis parts (1) of the electrode of arranging, and the monitoring component (2) of the big molecule electrophoretic band of monitoring bio position and the control circuit parts (3) of control electrode switch constitute; Wherein the electrophoresis parts comprise a main separating pipe and one or more lateral, and electrode is all arranged in the Xiao Chi of each duct end; Electrode contacts with buffer solution among the Xiao Chi by inserted mode, or by sputter or electroplating processing process be fixed in the Xiao Chi bottom directly and the buffer solution among the Xiao Chi contact.
4. according to the described biological macromolecule purifying retracting device of claim 3, it is characterized in that described electrophoresis parts are three-decker: it is the upper strata cover plate (6) that is processed with the pipe layers (5) of the electrophoresis pipeline of branch shape and cavity and is processed with corresponding aperture that the substrate that is processed with the electrode of arranging is positioned at bottom (4), intermediate layer.
5. according to the described biological macromolecule purifying retracting device of claim 4, it is characterized in that the zone line of bottom substrate (4) arranges the keeper electrode (9) of application of sample electrode (7), band shaping electrode (8), recovery sample, passive electrode (10) and main separate mesh electrode (12) successively; The periphery be being connected of drawing of each electrode with control circuit be connected electrode (11).
6. according to the described biological macromolecule purifying retracting device of claim 4, it is characterized in that: in the pipe layers that is processed with branch shape electrophoresis pipeline and cavity, with reference to electrophoresis pipeline (15) is single straight pipeline, and two ends have addition pool (13) and waste liquid pool (17) respectively; Purifying reclaims electrophoresis pipeline (14) than with reference to how one or more lateral of electrophoresis pipeline (15) and collecting pit.
7. according to the described biological macromolecule purifying retracting device of claim 4, it is characterized in that described cover plate layer comprises the well of application of sample and sampling (18), collection hole (19) and waste liquid hole (20); Well (18) position is corresponding to the addition pool (13) in the pipe layers (5), and collection hole (19) is corresponding to lateral and collecting pit (16) on the purifying recovery electrophoresis pipeline (14), and waste liquid hole (20) are corresponding to the waste liquid pool (17) on the pipe layers (3).
8. according to the described biological macromolecule purifying retracting device of claim 4, it is characterized in that: be positioned at the electrode of collecting pit or outside and coat one deck pellicle, perhaps and isolate the material of one deck pellicle between the lateral; Described pellicle only can penetrating small-molecule substance.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100242571A CN1314476C (en) | 2005-03-09 | 2005-03-09 | Method and equipment for purifying and recovering biomacromolecules |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005100242571A CN1314476C (en) | 2005-03-09 | 2005-03-09 | Method and equipment for purifying and recovering biomacromolecules |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1695775A CN1695775A (en) | 2005-11-16 |
| CN1314476C true CN1314476C (en) | 2007-05-09 |
Family
ID=35348762
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005100242571A Expired - Fee Related CN1314476C (en) | 2005-03-09 | 2005-03-09 | Method and equipment for purifying and recovering biomacromolecules |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN1314476C (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| BR112013032013A2 (en) * | 2011-06-16 | 2016-12-20 | British Columbia Cancer Agency | nucleic acid size selection method, nucleotide size selection apparatus and cassette for use in nucleic acid electrophoresis |
| WO2019237205A1 (en) * | 2018-06-14 | 2019-12-19 | Coastal Genomics Inc. | Device for capturing macromolecules and methods for manufacturing and using same |
| CN109456882A (en) * | 2019-01-16 | 2019-03-12 | 北京理工大学 | A kind of space biological sample micro process integrating device |
| JP7373923B2 (en) * | 2019-06-20 | 2023-11-06 | 株式会社日立ハイテク | Biological material recovery method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020187503A1 (en) * | 2001-05-02 | 2002-12-12 | Michael Harrold | Concentration and purification of analytes using electric fields |
| CN1395096A (en) * | 2001-07-11 | 2003-02-05 | 中国科学院大连化学物理研究所 | Stereo multi-dimensinal multi-mode capillary electrophoresis method and its special equipment |
| CN1469123A (en) * | 2002-07-18 | 2004-01-21 | 中国科学院大连化学物理研究所 | Microflow control chip for protein analysis and its application in protein analysis |
| WO2004051232A1 (en) * | 2002-11-29 | 2004-06-17 | Nec Corporation | Separating apparatus, separating method, and mass analyzing system |
-
2005
- 2005-03-09 CN CNB2005100242571A patent/CN1314476C/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020187503A1 (en) * | 2001-05-02 | 2002-12-12 | Michael Harrold | Concentration and purification of analytes using electric fields |
| CN1395096A (en) * | 2001-07-11 | 2003-02-05 | 中国科学院大连化学物理研究所 | Stereo multi-dimensinal multi-mode capillary electrophoresis method and its special equipment |
| CN1469123A (en) * | 2002-07-18 | 2004-01-21 | 中国科学院大连化学物理研究所 | Microflow control chip for protein analysis and its application in protein analysis |
| WO2004051232A1 (en) * | 2002-11-29 | 2004-06-17 | Nec Corporation | Separating apparatus, separating method, and mass analyzing system |
Non-Patent Citations (2)
| Title |
|---|
| 毛细管电泳与电泳芯片检测方法的研究 陈继峰，金庆辉，赵建龙，化学世界，第2期 2002 * |
| 电泳芯片系统的研制和应用 金庆辉，陈继峰，赵建龙，功能材料与器件学报，第7卷第4期 2001 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1695775A (en) | 2005-11-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0891369B1 (en) | Process and device for isolating nucleic acids | |
| KR100792683B1 (en) | - An Apparatus for Protein Separation Using Capillary Isoelectric Focusing-Hollow Fiber Flow Field Flow Fractionation and Method Thereof | |
| CN1314476C (en) | Method and equipment for purifying and recovering biomacromolecules | |
| CN102492603A (en) | Automated instrument for nucleic acid extraction and mass spectrum sample application | |
| DE102006050871B4 (en) | Integrated microfluidic component for the purification of analyte molecules as well as methods for purification | |
| JP7109596B2 (en) | Methods and systems for LC-MS/MS proteomic genotyping | |
| RU2011147219A (en) | METHOD AND DEVICE FOR ISOLATING THE Searched BIOPARTICLE FROM THE BIOLOGICAL SAMPLE | |
| WO2016112694A1 (en) | Device for nucleic acid extraction using magnetic bead method | |
| WO2006130632A3 (en) | Separation and purification of nucleic acid from paraffin-containing samples | |
| RU2004104953A (en) | METHOD FOR ISOLATING THE TETROMERIC FORM OF URICASE AND URICASE OBTAINED BY THIS METHOD | |
| Crowell et al. | A two-stage spin cartridge for integrated protein precipitation, digestion and SDS removal in a comparative bottom-up proteomics workflow | |
| JPH02504224A (en) | Method and apparatus for purifying M-13 fuzzy DNA | |
| CN1355319A (en) | Process for separating nucleic acid from biological particles by solid-phase carrier | |
| EP3988940A1 (en) | Full-automatic biological evaluation and chemical analysis integrated machine and method | |
| CN110592185A (en) | Method for designing hypercholesteremia virulence gene screening probe and gene chip thereof | |
| CN109053877B (en) | Method for extracting apolipoprotein B from serum and apolipoprotein B | |
| Martinez-Duarte | Carbon-electrode dielectrophoresis for bioparticle manipulation | |
| WO2004048398A1 (en) | Method and apparatus for concentration and purification of nucleic acid | |
| CA2535761A1 (en) | Parallel process for protein or virus separation from a sample | |
| EP1421360A1 (en) | Microscale affinity purification system | |
| CN110904095A (en) | Method for improving small fragment nucleic acid purification yield | |
| CN100350247C (en) | Method for differentiation and enrichment of cell specific target structure | |
| CN117051481B (en) | Preparation method and application of space bar code chip | |
| CN209481658U (en) | A kind of space biological sample micro process integrating device | |
| CN1277204A (en) | Nucleic acid purifying rotating centrifugal column and its usage in extracting nucleic acid |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| ASS | Succession or assignment of patent right |
Owner name: NINGBO RUIXIN BIOLOGY SCIENCE CO., LTD. Free format text: FORMER OWNER: SHANGHAI INST. OF MICROSYSTEM AND INFORMATION TECHNOLOGY, CHINESE ACADEMY OF SCI Effective date: 20080919 |
|
| C41 | Transfer of patent application or patent right or utility model | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20080919 Address after: Room 301, Pioneer Building, academician Road, Ningbo science and Technology Park, Zhejiang Patentee after: Ningbo realchip Biotechnology Co. Ltd. Address before: No. 865, Changning Road, Shanghai, Changning District Patentee before: Shanghai Institute of Microsystem and Information Technology, Chinese Academy of Sciences |
|
| C17 | Cessation of patent right | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20070509 |