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Crystallizer used for biomacromolecule crystallization, method and application thereof

A biomacromolecule and crystallization device technology, which is applied in the field of biomacromolecule crystallization devices, can solve the problems of amorphous precipitation and the inability to determine the crystallization concentration conditions of biomacromolecules, so as to reduce the amount of reagents and consumables, reduce costs, and expand the scope Effect

Inactive Publication Date: 2011-10-19
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the prior art crystallization method of biological macromolecules, the form of liquid-liquid mixing is adopted. In this form, the mixing speed of the two solutions is too fast. Once the concentration is too high, amorphous precipitation will appear immediately and it is impossible to determine the biological Concentration Conditions for Macromolecular Crystallization

Method used

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  • Crystallizer used for biomacromolecule crystallization, method and application thereof
  • Crystallizer used for biomacromolecule crystallization, method and application thereof
  • Crystallizer used for biomacromolecule crystallization, method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0072] choose as image 3 As shown in the crystallization plate, the crystallization plate has crystallization units 20 distributed in a 6×8 array, and each crystallization unit 20 has two titration areas 210 and one pool liquid area 220 . The screening solution is added to a titration area 210 of each crystallization unit 20 and then the crystallization plate is left to dry naturally at room temperature, that is, the screening solution is dried. Solid matter corresponding to the components of the screening fluid remains in the titration region 210 after drying. Into another titration area 210 in each crystallization unit 20, the same screening liquid corresponding to the previous operation is added again, but no drying treatment is required. Into the pool liquid area 220 of the crystallization unit 20, the same concentration but 50 times the screening liquid was also added.

[0073] Add the protein solution HutG to be crystallized into the crystallization area 210 in the cr...

Embodiment 2

[0098] The crystallization plates distributed in the form of a 6x8 array in which the pool liquid area and the crystallization area overlap were selected.

[0099] Screening solution:

[0100] Row A: 6% NaCl, 0.1M NaAc-HCl pH4.5

[0101] Row B: 7.5% NaCl, 0.1M NaAc-HCl pH4.5

[0102] Row C: 9% NaCl, 0.1M NaAc-HCl pH4.5

[0103] Row D: 0.02M calcium chloride dihydrate, 0.1M sodium acetate trihydrate pH4.6, 30% v / v (+ / -)-2-methyl-2,4-pentanediol

[0104] Row E: 0.2M Lithium Sulfate Monohydrate, 0.1M Tris Hydrochloride pH 8.5, 30% w / v Polyethylene Glycol 4000

[0105] Row F: 1.0M Na / K tartrate, 0.1M sodium 4-hydroxyethylpiperazineethanesulfonate pH7.5

[0106] Row G: 1.5M sodium formate, sodium acetate pH4.6

[0107]Row H: 1.0M Sodium Citrate, Sodium 4-Hydroxyethylpiperazineethanesulfonate pH7.5

[0108] crystallization process

[0109] Pre-drying:

[0110] 1 microliter of screening solution was added to each crystallization area 20, and the entire crystallization plate w...

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Abstract

The present invention relates to a crystallizer used for biomacromolecule crystallization. The crystallizer comprises at least one crystallizing unit, wherein the crystallizing unit includes at least The present invention relates to a crystallizer used for biomacromolecule crystallization. The crystallizer comprises at least one crystallizing unit, wherein the crystallizing unit includes at leastone pit liquid area for dripping pit liquid and at least one crystallizing area for dripping filtering liquid (commonly small amount of pit liquid) and the biomacromolecule liquid to be crystallized.one pit liquid area for dripping pit liquid and at least one crystallizing area for dripping filtering liquid (commonly small amount of pit liquid) and the biomacromolecule liquid to be crystallized.The crystallizing area is communicated with the pit liquid area. In at least one crystallizing area, the solid matter of the dried filtered liquid is reserved. The invention also relates to a method uThe crystallizing area is communicated with the pit liquid area. In at least one crystallizing area, the solid matter of the dried filtered liquid is reserved. The invention also relates to a method used for crystallizing the biomacromolecule. The method for crystallizing the biomacromolecule according to the invention can effectively control the crystallizing speed of biomacromolecule and effectised for crystallizing the biomacromolecule. The method for crystallizing the biomacromolecule according to the invention can effectively control the crystallizing speed of biomacromolecule and effectively confirming the conditions of concentration, etc. required by the crystallization of biomacromolecule thereby simplifying the crystallization experiment process of the biomacromolecule. Furthermorvely confirming the conditions of concentration, etc. required by the crystallization of biomacromolecule thereby simplifying the crystallization experiment process of the biomacromolecule. Furthermore the non-volatile solid ''filtering liquid'' is provided in the crystallizing area, therefore the crystallizer with the ''filtering liquid'' can be saved for a long time and can be used directly whene the non-volatile solid ''filtering liquid'' is provided in the crystallizing area, therefore the crystallizer with the ''filtering liquid'' can be saved for a long time and can be used directly when required.required.

Description

technical field [0001] The invention relates to a device for crystallization of biomacromolecules, a crystallization method for diffusing biomacromolecules solution by using dried screening liquid as an additive and its application. Background technique [0002] The determination of the structure of biological macromolecules is closely related to the understanding of the functions of biological macromolecules, the relationship with diseases and drug design. At present, X-ray crystallography is the main research method for the structure of biological macromolecules, which can be used for any crystallizable biological macromolecules, so that the crystallization technology of biological macromolecules becomes the focus of the research on the structure of biological macromolecules. [0003] The two main difficulties in analyzing the structure of biomacromolecules by X-ray crystallography are: providing sufficient samples of biomacromolecules and obtaining well-grown diffractive ...

Claims

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Application Information

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IPC IPC(8): B01D9/02B01L3/06C30B29/58
CPCC30B29/58B01D9/00C07K1/306C30B7/02
Inventor 苏晓东埃里克·布罗斯特罗梅尔
Owner PEKING UNIV