Preparation process for A beta oligomer antibody

A preparation process, oligomer technology, applied in the preparation method of peptides, anti-animal/human immunoglobulin, organic chemistry, etc., can solve the problems of low recovery rate of purification process, gel is not pressure-resistant, low flow rate, etc. , achieve the effect of high flow rate and ensure safety

Active Publication Date: 2017-03-22
SHANDONG TAIBANG BIOLOGICAL PROD CO LTD +1
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  • Summary
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  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although there are reports in the literature that affinity gels are used to extract and purify Aβ oligomer antibodies from IVIG, they are limited to laboratory research. The gels used are not pressure-resistant (maximum withstand pressure 0.015MPa), and the flow rate is low (maximum linear velocity 25cm / h ), the recovery rate of the purification process is low, and it is not suitable for industrialized production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Extract high concentration of Aβ oligomer antibody from intravenous human immunoglobulin (IVIG)

[0031] 1) Adjust the conductivity, pH value and protein concentration of the sample before loading the chromatography column

[0032] The 5L IVIG product (5%, 2.5g) was adjusted with 0.15mol / L sodium hydroxide to adjust the pH of the sample to 7.0, the conductivity was adjusted to 25mS / cm with the chromatography balance solution, the sample protein concentration was 8g / L, Aβ oligomer antibody The purity is 0.2%.

[0033] 2) Affinity chromatography

[0034] Pass the adjusted sample solution through the balanced GE Healthcare Blue gel affinity chromatography column. Wash the non-specifically adsorbed antibody in the chromatography column with the equilibration solution, and elute the adsorbed Aβ antibody with the eluent. The eluate was neutralized with 1.0 mol / L glycine (pH 2.0), and the pH was adjusted to 4.0. The sample load is 20 bed volumes.

[0035] Balance solution: 20mmol / L...

Embodiment 2

[0042] Extract high concentration of Aβ oligomer antibody from plasma component I+II+III precipitation

[0043] 1) Prepare IgG-rich feed solution from component I+II+III precipitation

[0044] Take 5000g of plasma fraction I+II+III precipitation, dissolve it with 50L 0.02mol / L acetic acid / sodium acetate buffer (pH4.5), after dissolution, use 0.3mol / L NaOH solution to adjust the pH to 4.5. After stirring for 1 hour, slowly add n-octanoic acid until the final concentration of n-octanoic acid in the solution is 0.11 mol / L, stop, and stir quickly for 2 hours to precipitate impurities; the solution is filtered through a step of ion exchange chromatography to obtain a feed solution rich in IgG.

[0045] 2) Adjust the conductivity, pH value and protein concentration of the sample before applying the chromatography column.

[0046] Adjust the pH of the IgG-rich feed solution obtained in step 1) to 6.5 with 0.15 mol / L sodium hydroxide, adjust the conductivity to 50 mS / cm with the chromatograph...

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Abstract

The invention discloses a preparation process for an A beta oligomer antibody. The preparation process comprises the following steps that the conductivity, the pH and the protein concentration of an upper chromatographic column front sample are adjusted; affinity gel chromatography is conducted; eluent from an affinity chromatographic column is filtered, ultra-filtered and prepared; and virus inactivation and sterilized sub-packaging are conducted. According to the preparation process for the A beta oligomer antibody, one-step affinity chromatography is adopted, the purification multiple is 85-100 times, and the A beta oligomer antibody obtained through purification is high in concentration and clear in targeting performance and has wide clinical application prospects.

Description

Technical field [0001] The present invention relates to the technical field of antibody preparation, in particular to a preparation process of Aβ oligomer antibody. Background technique [0002] Alzheimer’s disease (AD), also known as senile dementia, was first discovered by German doctor Alois Alzheimer in 1907 and is the most common central nervous system degenerative disease. The main symptoms of AD are neuropsychiatric symptoms such as progressive memory and cognitive dysfunction and language disorders. As the country’s population becomes more aging, the social and economic problems caused by AD will become increasingly severe. [0003] There are many theories on the pathogenesis of AD, among which the "amyloid β-protein (Aβ) theory" is widely accepted as the main pathogenesis of AD. The molecular weight of Aβ is about 4kDa, which is hydrolyzed from β-amyloid precursor protein (APP). Aβ can aggregate to form Aβ oligomers with toxic effects, leading to the occurrence and deve...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/18C07K1/36C07K1/34C07K1/22
CPCC07K1/22C07K1/34C07K1/36C07K16/18
Inventor 宋佃卫谢喜秀马山刘瑞田菅长永朱孟沼席智赢刘坤范克明
Owner SHANDONG TAIBANG BIOLOGICAL PROD CO LTD
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