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Isoelectric point standard substance for free flow electrophoresis

An isoelectric point and standard product technology, applied in the field of free-flow electrophoresis, can solve the problems that it is difficult to meet the requirements of different systems, and the poor effect of positioning and separating protein drug charge isomers, so as to shorten the preparation time and improve the positioning effect.

Pending Publication Date: 2021-11-02
上海蓝析生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Isoelectric point standards with different colors can accurately locate the positions of different protein drug charge variants during separation, but the existing isoelectric point standards are not effective in positioning and separating protein drug charge variants. Difficult to meet the requirements of different systems

Method used

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  • Isoelectric point standard substance for free flow electrophoresis
  • Isoelectric point standard substance for free flow electrophoresis
  • Isoelectric point standard substance for free flow electrophoresis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] An isoelectric point standard product for free-flow electrophoresis, comprising: isoelectric point substances, the isoelectric point substances are provided with 8 kinds, and the isoelectric point substances are composed of small molecular substances diluted with ultrapure water, so The molecular weight of the isoelectric point substances is between 200-600KDa, all of which are benzene rings and heterocyclic substances, and the isoelectric point substances are respectively 4,6-diaminoazobenzene-2,2-dicarboxylic acid, 3 ', 3"-bis[N-(4-carboxypiperazinyl)methyl]-o-cresolsulfonphthalein, 3',5',3"5"-tetrakis(N-[4-carboxypiperazine]methyl Base)-phenolsulfophthalic acid, 3',3"-bis(4-morpholinomethyl)-5',5"-dichlorophenolsulfonphthalein, 3',3"-bis(4-morpho Phenylmethyl)-o-cresolsulfonphthalein, 2,4-bis-(4-morpholinomethyl)-6-nitrophenol, 2,4-bis[(4-methyl-1-piperazinyl) Methyl]-6-nitrophenol, 2-nitro-4,6-bis(1-piperidylmethyl)phenol, and each isoelectric point substance corre...

Embodiment 2

[0023] An isoelectric point standard product for free-flow electrophoresis, comprising: isoelectric point substances, the isoelectric point substances are provided with 8 kinds, and the isoelectric point substances are composed of small molecular substances diluted with ultrapure water, so The molecular weight of the isoelectric point substances is between 200-600KDa, all of which are benzene rings and heterocyclic substances, and the isoelectric point substances are respectively 4,6-diaminoazobenzene-2,2-dicarboxylic acid, 3 ', 3"-bis[N-(4-carboxypiperazinyl)methyl]-o-cresolsulfonphthalein, 3',5',3"5"-tetrakis(N-[4-carboxypiperazine]methyl Base)-phenolsulfophthalic acid, 3',3"-bis(4-morpholinomethyl)-5',5"-dichlorophenolsulfonphthalein, 3',3"-bis(4-morpho Phenylmethyl)-o-cresolsulfonphthalein, 2,4-bis-(4-morpholinomethyl)-6-nitrophenol, 2,4-bis[(4-methyl-1-piperazinyl) Methyl]-6-nitrophenol, 2-nitro-4,6-bis(1-piperidylmethyl)phenol, and each isoelectric point substance corre...

Embodiment 3

[0030] An isoelectric point standard product for free-flow electrophoresis, comprising: isoelectric point substances, the isoelectric point substances are provided with 8 kinds, and the isoelectric point substances are composed of small molecular substances diluted with ultrapure water, so The molecular weight of the isoelectric point substances is between 200-600KDa, all of which are benzene rings and heterocyclic substances, and the isoelectric point substances are respectively 4,6-diaminoazobenzene-2,2-dicarboxylic acid, 3 ', 3"-bis[N-(4-carboxypiperazinyl)methyl]-o-cresolsulfonphthalein, 3',5',3"5"-tetrakis(N-[4-carboxypiperazine]methyl Base)-phenolsulfophthalic acid, 3',3"-bis(4-morpholinomethyl)-5',5"-dichlorophenolsulfonphthalein, 3',3"-bis(4-morpho Phenylmethyl)-o-cresolsulfonphthalein, 2,4-bis-(4-morpholinomethyl)-6-nitrophenol, 2,4-bis[(4-methyl-1-piperazinyl) Methyl]-6-nitrophenol, 2-nitro-4,6-bis(1-piperidylmethyl)phenol, and each isoelectric point substance corre...

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Abstract

The present invention provides an isoelectric point standard substance for free flow electrophoresis, the isoelectric point standard substance comprises eight isoelectric point substances, each isoelectric point substance corresponds to a different isoelectric point, each isoelectric point substance has a color capable of being directly observed by naked eyes, the isoelectric point of each isoelectric point substance is between 3.8 and 10.1, the isoelectric point substance is composed of micromolecular substances diluted by ultrapure water, the molecular weight of the isoelectric point substances is 200-600 KDa, and the isoelectric point substances are benzene ring and heterocyclic ring substances. The isoelectric point standard substance meets the system standards of pH 3-10, pH 7-10 and pH 5-8, the pH can be positioned in the isomer separation and preparation process, the isomer positioning is greatly improved, and the isomer preparation time is shortened.

Description

technical field [0001] The invention belongs to the technical field of free-flow electrophoresis, and in particular relates to an isoelectric point standard product for free-flow electrophoresis. Background technique [0002] Charge variant analysis is a regulatory requirement for biotherapeutic proteins. These large heterogeneous molecules undergo various enzymatic post-translational modifications during production, such as glycosylation and lysine cleavage. In addition, various chemical modifications such as oxidation or deamination can occur during purification and storage. Free-flow electrophoresis is the best method for preparing protein drug charge variants. [0003] Free Flow Electrophoresis (FFE) is a semi-preparative separation technology, which has the advantages of continuous separation, multiple separation modes, no fixed support medium and mild separation conditions, and is especially suitable for the separation, purification and preparation of biological mate...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28G01N27/447
CPCG01N1/28G01N27/447
Inventor 马艳师要辉吴清灵马校卫张思齐沈治峰袁新玲
Owner 上海蓝析生物技术有限公司
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