Chromatographic medium for capturing mRNA and preparation method thereof

A chromatographic medium and medium technology, applied in the field of chromatographic medium and its preparation, can solve the problems of unsatisfactory removal of impurities and unfavorable large-scale purification of mRNA products, etc., and achieve good rigidity, high capacity, and good surface hydrophilicity Effect

Pending Publication Date: 2021-12-17
SEPAX TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among these products, due to limited principles, magnetic beads can only be used in laboratory research to purify a small amount of mRNA; cellulose is not conducive to large-scale downstream purification of mRNA products because the matrix is ​​too soft; polyethylene-divinylbenzene microspheres Good rigidity, can be used for industrial purification of mRNA products, but due to its strong surface hydrophobicity, other impurities can be bound to the microspheres through hydrophobic interaction, mixed into mRNA products in the subsequent elution steps, and the removal effect of impurities is not ideal

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] The basic medium used in this example is polymethacrylate microspheres, the product name is Monomix MC30-SEC-1000 (article number: 280130950), and the specifications are: particle size 30m monodisperse, pore size The microspheres are hydrophilically modified and produced by Suzhou Saifen Technology Co., Ltd. The rest of the raw materials are commercially available, and the base length of Oligo dT is 20.

[0031] [S01]-001

[0032] Take 5L Monomix MC30-SEC-1000 in a 20L reaction kettle with a measuring cylinder, add 5L 1mol / L sodium hydroxide solution and turn on the mechanical stirring to 50rpm, after running for 10min, add 5L 1,4-butanediol diglycidyl ether at one time , the temperature was raised to 45°C and the reaction was continuously stirred for 4 hours. After the end, the medium was transferred to a Buchner funnel and filtered with suction, then washed 5 times with 5L of absolute ethanol, and washed with water until the filtrate was neutral.

[0033] [S02]-001 ...

Embodiment 2

[0039] The basic medium used in this example is polymethacrylate microspheres, the product name is Monomix MC60-SEC-1000 (article number: 280160950), and the specifications are: particle size 60m monodisperse, pore size The microspheres are hydrophilically modified and produced by Suzhou Saifen Technology Co., Ltd. The rest of the raw materials are commercially available, and the base length of Oligo dT is 30.

[0040] [S01]-002

[0041] Take 2.5L Monomix MC60-SEC-1000 into a 10L reaction kettle with a graduated cylinder, add 2.5L 1.2mol / L sodium hydroxide solution and turn on the mechanical stirring to 50rpm, after running for 10min, add 2.5L 1,6-hexanediol at one time For diglycidyl ether, the temperature was raised to 35°C and the reaction was continuously stirred for 3 hours. After the end, the medium was transferred to a Buchner funnel and filtered with suction, then washed 5 times with 2.5L absolute ethanol, and washed with water until the filtrate was neutral.

[0042...

Embodiment 3

[0048] The basic medium used in this example is polymethacrylate microspheres, the product name is Monomix MC30-SEC-1000 (article number: 280130950), and the specifications are: particle size 30m monodisperse, pore size The microspheres are hydrophilically modified and produced by Suzhou Saifen Technology Co., Ltd. The rest of the raw materials are commercially available, and the base length of Oligo dT is 15.

[0049] [S01]-003

[0050] Take 500mL Monomix MC30-SEC-1000 in a 2L reaction kettle with a measuring cylinder, add 500mL 0.3mol / L sodium hydroxide solution and turn on the mechanical stirring to 50rpm, and add 500mL 1,2-ethylene glycol diglycidyl at one time after running for 10min Ether, the temperature was raised to 37°C and the reaction was continuously stirred for 5 hours. After the end, the medium was transferred to a Buchner funnel and filtered with suction, then washed 5 times with 5L of absolute ethanol, and washed with water until the filtrate was neutral.

...

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Abstract

The invention discloses a chromatography medium for capturing mRNA and a preparation method thereof, the structure is SP-R-Oligo dT, SP is a basic medium, Oligo dT is an affinity ligand, and R is a spacer arm of the basic medium coupled Oligo dT. The method comprises the following steps: (1) activating a basic medium to obtain an activated medium with a large number of epoxy groups or aldehyde groups modified on the surface; (2) coupling the Oligo dT to an activation medium; and (3) carrying out sealing treatment on redundant aldehyde groups or epoxy groups to obtain the Oligo dT affinity medium. The medium is good in rigidity, good in surface hydrophilicity, high in loading capacity and suitable for downstream purification of mRNA prevention and treatment biological products.

Description

technical field [0001] The invention relates to a chromatographic medium and a preparation method thereof, in particular to a chromatographic medium for capturing mRNA and a preparation method thereof. Background technique [0002] As early as the 1990s, scientists injected mRNA synthesized in vitro into mice, and found that it could be translated into related active proteins in mice, and produced an immune response, which could play the role of a vaccine. This is The prototype of the mRNA vaccine. However, due to technical limitations, mRNA has many bottlenecks in terms of stability, safety and drug delivery. After 30 years of development, mRNA synthesis, modification, and delivery technology have made great progress, especially the pandemic of the new coronavirus, which prompted the first mRNA vaccine in human history to be approved for disease prevention. In terms of effectiveness, mRNA vaccines It is significantly higher than that of traditional technology vaccines, wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/285B01J20/286C12N15/10B01D15/38B01J20/30
CPCB01J20/285B01J20/286B01D15/3804C12N15/101
Inventor 吕小林丁良龙陈迪安胡新妹毛慧明
Owner SEPAX TECH
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