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Method to promote protein crystallization with bioglass

A technology of protein crystallization and biological glass, which is applied to the preparation method of peptides, chemical instruments and methods, organic chemistry, etc., can solve the problems of loss, large particles, inconvenient and other problems, and achieve uniform dispersion, strong surface binding, and non-toxicity. easy precipitation effect

Inactive Publication Date: 2015-08-19
江苏同庚电子科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] When the above-mentioned technical solution uses mesoporous glass (that is, the above-mentioned bioglass), the glass is directly put into the protein solution to promote protein crystallization; Leave it for a period of time to observe the crystallization of the protein; since the bioglass particles are very small, use tweezers to take the bioglass, and it is not very convenient to take it out with tweezers; It loses its effect; and in order to match the way of tweezing, the particles of bioglass are larger, so the holes inside the bioglass also lose their effect

Method used

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  • Method to promote protein crystallization with bioglass
  • Method to promote protein crystallization with bioglass
  • Method to promote protein crystallization with bioglass

Examples

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Embodiment 1

[0033] (Example 1, the method of promoting protein crystallization with bioglass)

[0034] The bioglass suspension in this embodiment is a mixture formed by suspending small bioglass particles in deionized water. The small bioglass particles are ultrafine bioglass particles with a particle size of less than 200 nm. In the bioglass suspension, the concentration of the bioglass small particles is 2-50 / μL, and the concentration in this embodiment is 30 / μL.

[0035] The method that biological glass suspension impels protein crystallization comprises the steps:

[0036] ①Preparation of bioglass: P 2 o 5 :CaO:MgO:SiO 2 =1:3:1:5, weigh the corresponding P 2 o 5 , Ca(NO 3 ) 2 , Mg(NO3) 2 , tetraethyl orthosilicate, and then each compound was mixed with absolute ethanol and heated to reflux to dissolve in ethanol to obtain uniform P 2 o 5 ethanol solution, Ca(NO 3 ) 2 ethanol solution, Mg(NO3) 2 The ethanol solution and the ethanol solution of ethyl orthosilicate; the mol...

Embodiment 2

[0049] (Example 2, the method of promoting protein crystallization with bioglass)

[0050] In this example, the rest of the biological glass suspension obtained in step ② is the same as in Example 1, except that the suspension also includes 20 mg / mL of chloroform, 2.5 mg / mL of the porogen polyethylene glycol Alcohol 3350 and 5 mg / mL active sulfated oil. That is to say, the liquid solvent for increasing the surface activity of bioglass in this embodiment is deionized water, and the solute includes chloroform with a concentration of 20 mg / mL, 2.5 mg / mL porogen polyethylene glycol 3350 and 5 mg / mL active agent sulfated oil . The sulfated oil used in this embodiment is the sulfonated oil obtained by the action of castor oil and sulfuric acid, and the molecular formula is C 18 Na 2 o 6 S, CAS Registry No. 8002-33-3.

[0051] The biological glass suspension of the present embodiment promotes the remainder of the method for protein crystallization to be the same as in Example 1,...

Embodiment 3

[0055] (Example 3, the method for promoting protein crystallization with bioglass)

[0056] The rest of this embodiment is the same as embodiment 2, the difference is:

[0057] In step ③, when the protein crystallization is promoted by the bioglass suspension, the bioglass suspension promotes protein crystallization in the microfluidic protein crystallization plate.

[0058] See figure 2 , the microfluidic protein crystallization plate used in this embodiment is an injection-molded integral piece, including a support structure part 1 and crystallization units 2 connected to the support structure part 1 and distributed in an array.

[0059] The supporting structure part 1 is composed of a panel 11 and a supporting part 12 supported on the periphery of the panel 11 . The overall shape of the panel 11 is a rectangle with one corner missing, and the triangular notch is used as a mark of the placement position during use, and the panel 11 is provided with a cross-sectional shape...

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Abstract

The invention discloses a method for prompting protein crystallization by the use of bioglass. According to the method, superfine bioglass particles are mixed with a liquid for increasing surface activity of bioglass to form a suspension with long service life and stable properties; and the suspension is mixed with a protein saturated solution so as to form an environment which is beneficial to crystallization. After bioglass is made into the suspension, bioglass can be dripped by the use of a dropper to a place where the bioglass is required. Therefore, an application method of bioglass is greatly improved, and automation can replace manual work. The method is more suitable for large-scale protein crystallization condition screening. As the suspension, bioglass can be fully mixed with a protein solution when in use, and the contact surface with protein is larger, which is beneficial to crystallization under various conditions.

Description

technical field [0001] The invention relates to the field of protein crystallization, in particular to a method for promoting protein crystallization by using biological glass. Background technique [0002] With the development of biotechnology, people pay more and more attention to the unique functional properties of proteins and other biomacromolecules. Among them, the medicinal properties of many proteins are constantly being discovered. As we all know, the functional characteristics of a molecule mainly depend on its unique three-dimensional space structure. Therefore, to understand and determine the function of a certain biomacromolecule, it is necessary to measure and study its three-dimensional structure; It is of great significance to change or modify the molecular structure and make it have specific application properties. [0003] Currently, there are three main methods for protein structure determination: X-ray diffraction, nuclear magnetic resonance and electron...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K1/14
Inventor 李云胡春良彭萍
Owner 江苏同庚电子科技有限公司
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