CN203229482U - Novel protein crystallization device - Google Patents
Novel protein crystallization device Download PDFInfo
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- CN203229482U CN203229482U CN 201320245713 CN201320245713U CN203229482U CN 203229482 U CN203229482 U CN 203229482U CN 201320245713 CN201320245713 CN 201320245713 CN 201320245713 U CN201320245713 U CN 201320245713U CN 203229482 U CN203229482 U CN 203229482U
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Abstract
The utility model provides a novel protein crystallization device which comprises a crystallization plate and a transparent rubber belt, wherein a plurality of liquid storage slots are formed in the crystallization plate; a protein solution to be crystallized is put into the liquid storage slots; the transparent rubber belt of which the glue surface is provided with a drying agent array is stuck to the crystallization plate; after the transparent rubber belt is stuck, each drying agent on the transparent rubber belt corresponds to the position of each liquid storage slot on the crystallization plate; the drying agents are not in contact with the liquid storage slots, so that a sealed protein crystallization space is formed. The novel protein crystallization device can use the quickly produced compound drying agent array, save the labor, improve the production efficiency and improve the protein crystallization screening success rate and can be popularized and applied.
Description
Technical field
The present invention relates to a kind of protein crystallization apparatus, the device especially for improving the crystallization of protein success ratio belongs to the crystallization of protein field.
Background technology
The statistic data of drawing according to the structure gene batch total, from the very high protein of purity to the protein crystal that grows the diffraction quality, its success ratio less than 20%.The process that obtains protein crystal is to begin to a series of flow processs that obtain structure one of link that success ratio is minimum from expression.Therefore, the growing high-quality protein crystal is still one of the bottleneck problem in structure biology field.
The major reason that the crystallization of protein success ratio is low is that screening is less than the suitable crystallizing agent protein crystal of growing.The most frequently used crystallization of protein screening method is the gas phase diffusion method.The gas phase diffusion ratio juris is in a closed system, because the concentration of crystallizing agent is higher than the concentration of crystallizing agent in the crystallization of protein drop in the liquid of nonprotein pond, cause the constantly pond liquid diffusion from the crystallization drop of lower concentration to high density of solvent, the concentration of protein soln and crystallizing agent solution constantly increases in the crystallization drop thereby make, the protein drop reaches supersaturation gradually in this process, thereby forms crystal.In traditional gas phase diffusion method, crystallization drop concentration constantly concentrate until with pond liquid balance, if the concentration deficiency of pond liquid is so that crystallization drop is concentrated into the degree of supersaturation of protein forming core, even then employed screening reagent can obtain crystal under greater concn, but when screening, reality can not get crystal.
People such as deer celery celery utilize silica-gel drier to substitute pond liquid in the gas phase diffusion method, siccative plays the effect that highly concentrates drop, the concentration range of drop is significantly expanded, utilize the siccative method to carry out the screening of protein molecule crystallization condition, improved protein molecule crystallization screening success ratio (Replace reservoir solution with desiccant in vapor diffusion protein crystallization screening, Journal of Applied Crystallography, 2010, Vol.43:1021-1026), developed desiccating method step by step afterwards again: add silica-gel drier in batches, condensing crystal drop step by step, the concentration range of drop is significantly expanded until the drop evaporate to dryness, scan the more strength of solution of wide region with this, this method has further improved success ratio (the A gradual desiccation method for improving the efficiency of protein crystallization screening of crystallization of protein screening, Journal of Applied Crystallography, 2012, Vol.45:758-765).
But when applying step by step drying means, there is an obstacle, is the siccative that each crystallization drop prepares appropriate amount.Be example with 96 condition crystallization screening reagent boxes, each dry when protein is carried out crystallization screening, all need to control the amount of the siccative that 96 crystallization drop use.Therefore the primary crystallization drying process need be weighed 96 times at least, seeks silica-gel drier identical in quality, implements repeatedly when dry, at least will be to siccative weigh 96 * n time (n is dry number of times).In addition, each crystallization drop will with siccative need manual allocation in the deep hole of crystallization plates, this means needs other 96 * n time manual operation.All manual operations are repeatedly wasted time and energy.And, if the silica-gel drier particle is excessive, by crushing, weighing, be difficult to accurately control quality, simultaneously, the size of the silica gel particle that each crystallization drop is used is often variant, the dynamic process of lyosorption is different, and this can cause serious repeated problem.Therefore simultaneously, be not specifically designed to the crystallization plates of desiccating method step by step, have a deep hole to be used for specially adorning pond liquid in traditional crystallization plates, and desiccating method do not need pond liquid step by step, traditional crystallization plates is for desiccating method step by step and not exclusively be suitable for.
Summary of the invention
In order to overcome the deficiencies in the prior art, the invention provides a kind of protein crystallization apparatus, this device can utilize the compound siccative array of quick preparation, can save the labour, enhances productivity, and improves crystallization of protein and is filtered into power, applies.
The technical solution adopted for the present invention to solve the technical problems is: comprise crystallization plates and scotch tape.Described crystallization plates is provided with several reservoirs, place protein soln to be crystallized in the reservoir, to cover the scotch tape that the glue face is stained with the siccative array is bonded on the crystallization plates, paste the reservoir position of the respectively corresponding crystallization plates of every siccative on the scotch tape of back, and siccative does not contact mutually with reservoir, forms the crystallization of protein space of sealing.
The number of described reservoir is 96,192,288 or 384.
Described crystallization plates selects transparent material to make.
The invention has the beneficial effects as follows:
1, utilizes protein crystallization apparatus of the present invention, realize fast the purpose of drying protein crystallization solution step by step.
2, utilize protein crystallization apparatus of the present invention, compare with traditional gas phase diffusion method, in the crystallization of protein process, realize maximum range interscan protein soln concentration, widened crystallization of protein strength of solution interval, make crystallization solution reach supersaturation, do not miss any condition that can impel crystallization of protein, improved the success ratio of crystallization of protein screening greatly.
3, crystallization of protein of the present invention is filtered into power and is higher than traditional gas phase diffusion method.Compare with the method for protein crystallization of routine, adopt the inventive method to carry out the crystallization of protein screening, antalzyme crystallization is filtered into power and has been improved 30%, sweet protein crystallization screening success ratio has improved 60%, opalescin crystallization screening success ratio has improved 100%, Proteinase K crystallization screening success ratio has improved 80%, chymotrypsinogen A II crystallization screening success ratio has improved 120%, con A VI crystallization screening success ratio has improved 15%, ribonuclease A I crystallization screening success ratio has improved 8 times, and cytochrome c crystallization screening success ratio has improved 2 times.
4, the present invention is simple to operate, the efficient height, and the operating procedure condition is controlled easily, and is repeatable high, can mass production, realize applying.
Description of drawings
Fig. 1 is the perspective view of protein crystallization apparatus of the present invention;
Fig. 2 is the scotch tape that is stained with the solid drier array;
Fig. 3 is the vertical view of crystallization of protein plate of the present invention;
Fig. 4 is along the sectional view of A-A line among Fig. 3;
Among the figure: 1-crystallization plates body; The 2-reservoir; 3-is stained with the scotch tape of solid drier array.
Embodiment
The present invention is further described below in conjunction with drawings and Examples.
As shown in Figure 1, device of the present invention is by crystallization plates 1 and cover the scotch tape 3 that the glue face is stained with the siccative array and forms, wherein is provided with reservoir 2 at crystallization plates 1, is used for placement crystallization of protein solution, and 4 reservoirs are a little space.The scotch tape 3 that is stained with siccative is bonded on the crystallization plates 1, seals described reservoir 2, and every corresponding 4 reservoirs of siccative form the crystallization of protein space that seals.Compare with traditional crystallization plates, crystallization plates of the present invention does not have traditional method to be used to adorn the deep hole of pond liquid, realizes proteins concentrate crystallization solution step by step because the present invention utilizes siccative to replace pond liquid, has widened crystallization of protein strength of solution interval.
The number of reservoir of the present invention is 96,192,288, or 384,1,2,3,4 reservoirs namely can be set in each little space.
Crystallization plates of the present invention selects the material of transparent material to make;
The scotch tape that is stained with the siccative array of the present invention can be by two kinds of method preparations.1) the transparent sealing adhesive tape that covers glue at single face covers the glue face and prepares the siccative array: the preparation compound solution, utilize some means, and realize that solid compounds desiccant particle array is bonded at single face transparent sealing adhesive tape and covers the glue face.The indication means comprise two kinds: a. on glossy paper with solution according to moving liquid with the position of crystallization of protein plate reservoir coupling, be prepared into the drop array, the oven dry back obtains compound siccative array, closely stick on the glossy paper of siccative array with scotch tape again, tear scotch tape, namely cover the glue face at scotch tape and obtain the desiccants array.B. the no glue face that single face is covered the transparent sealing adhesive tape of glue with double faced adhesive tape sticks on the platform, covers glue and faces up.Select volume, distance and the array quantity of compound solution, with automatic liquor-transferring system or electronic pipettor or manually pipettor the adhesive tape that compound solution is ejected into above the platform is covered Jiao Mianshang, formation compound drop array.The adhesive plaster that is stained with the drop array is taken off from platform, put into the stoving oven oven dry, can realize that solid compounds desiccant particle array is bonded at adhesive tape and covers the glue face.
The present invention utilizes above-mentioned protein crystallization apparatus to carry out crystallization of protein to comprise following step in sequence:
1) preparation crystallization of protein solution according to demand, is prepared single or multiple crystallization of protein solution.
2) crystallization of protein: crystallization of protein solution is dripped in the reservoir of crystallization plates, with the sealant tape sealing crystallization plates that is stained with solid compounds siccative array, make the corresponding little space of containing 4 reservoirs of every siccative, and do not contact mutually, the crystallization plates temperature control under 0-55 ℃ of condition, is carried out crystallization, and the siccative array begins to draw the solvent in the crystallization solution, take out after 48-96 hour, observe the crystallization result.Each a slice siccative array of replacing repeats crystallization treatment, in confined conditions till the whole dryings of crystallization of protein solution.
It is the Index of HR2-144 that the present invention adopts Hampton company article No.
TM96 crystallization test kits and new crystallization apparatus be to N,O-Diacetylmuramidase, sweet protein, and opalescin, Proteinase K, chymotrypsinogen A II, con A VI, ribonuclease A I, cytochrome c has carried out crystallization screening.Except Hampton company article No. is the Index of HR2-144
TMOutside the 96 crystallization test kits, other test kits all are applicable to the present invention.Except N,O-Diacetylmuramidase, sweet protein, opalescin, Proteinase K, chymotrypsinogen A II, con A VI, ribonuclease A I, outside the cytochrome c, other protein all are applicable to the present invention.
Claims (3)
1. novel protein crystallization apparatus, comprise crystallization plates and scotch tape, it is characterized in that: described crystallization plates is provided with several reservoirs, place protein soln to be crystallized in the reservoir, to cover the scotch tape that the glue face is stained with the siccative array is bonded on the crystallization plates, paste the reservoir position of the respectively corresponding crystallization plates of every siccative on the scotch tape of back, and siccative does not contact the crystallization of protein space of formation sealing mutually with reservoir.
2. novel protein crystallization apparatus according to claim 1, it is characterized in that: the number of described reservoir is 96,192,288 or 384.
3. novel protein crystallization apparatus according to claim 1 is characterized in that: described crystallization plates selects transparent material to make.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201320245713 CN203229482U (en) | 2013-05-09 | 2013-05-09 | Novel protein crystallization device |
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| Application Number | Priority Date | Filing Date | Title |
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| CN 201320245713 CN203229482U (en) | 2013-05-09 | 2013-05-09 | Novel protein crystallization device |
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| CN 201320245713 Expired - Fee Related CN203229482U (en) | 2013-05-09 | 2013-05-09 | Novel protein crystallization device |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104330426A (en) * | 2014-11-06 | 2015-02-04 | 西北工业大学 | Device and method for screening crystallization condition of high-throughput protein under strong magnetic field |
| CN108137647A (en) * | 2015-10-09 | 2018-06-08 | 东丽株式会社 | Fibre-bearing crystal, the manufacturing method of fibre-bearing crystal, the manufacturing device of fibre-bearing crystal and chemical reagent infuser device |
| CN108159730A (en) * | 2017-12-27 | 2018-06-15 | 大连理工大学 | A kind of high throughput with the accurate continuously macromolecule crystal of micron scale construction prepares platform and method |
| CN108593689A (en) * | 2018-03-29 | 2018-09-28 | 西北工业大学 | A kind of diffraction device in situ and original position diffraction method of protein crystal |
-
2013
- 2013-05-09 CN CN 201320245713 patent/CN203229482U/en not_active Expired - Fee Related
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104330426A (en) * | 2014-11-06 | 2015-02-04 | 西北工业大学 | Device and method for screening crystallization condition of high-throughput protein under strong magnetic field |
| CN104330426B (en) * | 2014-11-06 | 2016-11-02 | 西北工业大学 | High throughput protein crystallization condition screening plant under a kind of high-intensity magnetic field and method |
| CN108137647A (en) * | 2015-10-09 | 2018-06-08 | 东丽株式会社 | Fibre-bearing crystal, the manufacturing method of fibre-bearing crystal, the manufacturing device of fibre-bearing crystal and chemical reagent infuser device |
| CN108159730A (en) * | 2017-12-27 | 2018-06-15 | 大连理工大学 | A kind of high throughput with the accurate continuously macromolecule crystal of micron scale construction prepares platform and method |
| CN108593689A (en) * | 2018-03-29 | 2018-09-28 | 西北工业大学 | A kind of diffraction device in situ and original position diffraction method of protein crystal |
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| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131009 Termination date: 20180509 |