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Monoclonal antibody purification process

A technology of monoclonal antibodies and process methods, applied in chemical instruments and methods, peptide preparation methods, organic chemistry, etc., can solve problems such as low resolution and poor separation effect, achieve improved purity, improved main peak purity, and improved antibody The effect of purity

Active Publication Date: 2015-11-04
上海药明生物医药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the resolution of the conventional elution mode is low, which can only slightly improve the purity of the target protein, and the separation effect of acid-base analogues with small charge differences from the target protein is poor

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1 Herceptin monoclonal antibody purification

[0024] 1. Protein A affinity chromatography

[0025] Step 1, wash the chromatography column with 3 times the column volume of water for injection, then sterilize the chromatography column with a mixed solution of 3 times the column volume of NaOH (50mM) and NaCl (1M), and then use 3 times the column volume of water for injection Rinse the column.

[0026] Step 2, equilibrate the chromatography column with 5 times the column volume of buffer solution (20mM disodium hydrogen phosphate-sodium dihydrogen phosphate + 150mM NaCl, pH7.4).

[0027] Step 3, sample loading, let the sample stay on the column for 4 minutes.

[0028] Step 4, wash with 20mM disodium hydrogen phosphate-sodium dihydrogen phosphate+150mM NaCl, pH7.4 to A280 baseline, and then use 5 times column volume buffer (20mM citric acid-sodium citrate+1M NaCl, pH5. 5) Wash the chromatographic column, and then wash the chromatographic column with 3 times ...

Embodiment 3

[0062] Embodiment 3 Rituximab monoclonal antibody purification

[0063] The purification process is the same as in Example 1, and the HCP and DNA residues and SEC purity of the affinity chromatography eluate after one-step deep filtration are shown in Table 6:

[0064] Table 6 HCP and DNA residues and SEC purity of affinity chromatography eluate after deep filtration

[0065]

[0066] It can be seen from Table 6 that after one-step deep filtration of the affinity chromatography eluate, the HCP residual content was significantly reduced, from 794ppm to 127ppm, and the DNA residue also fully complied with FDA requirements.

[0067] The purity and residue indicators of the final product are shown in Table 7:

[0068] Table 7 adopts the purity and residual index of the product prepared by process purification of the present invention

[0069]

[0070] It can be seen from Table 7 that the purity and residual indicators of the rituximab product purified and prepared by the p...

Embodiment 4

[0071] Example 4 MabThera Monoclonal Antibody Purification

[0072] The processing method of purification is the same as embodiment 1, and the purity and residual index of final product are referring to table 8:

[0073] Table 8 adopts the purity and residual index of the product prepared by process purification of the present invention

[0074]

[0075] It can be seen from Table 8 that the purity and residual indicators of the MabThera monoclonal antibody product purified and prepared by the process of the present invention also meet the requirements of sFDA.

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Abstract

The present invention discloses a monoclonal antibody purification process, which comprises: 1) carrying out affinity chromatography; 2) adjusting the pH value of the affinity chromatography eluent to 3.3-3.8, and carrying out virus inactivation; 3) adjusting the pH value to a neutral pH value, and carrying out deep filtration; 4) carrying out anion exchange chromatography; and 5) carrying out cation exchange chromatography. According to the present invention, the low-pH value and high-salt washing step is added to the affinity chromatography to remove the host protein and DNA while the deep filtration is used after the low-pH value virus inactivation to carry out the clarifying treatment, such that the separation effect of the affinity chromatography on the antibody monomer and the polymer is improved so as to increase the purity of the monoclonal antibody product.

Description

technical field [0001] The invention relates to the field of biology, in particular to a purification process of monoclonal antibodies. Background technique [0002] In 1997, the Genentech Corporation of the United States successfully developed the first humanized monoclonal antibody drug, taking the first step in the industrialization of monoclonal antibody drugs. In a short history of more than 10 years, antibody drugs have shown an unstoppable development trend. Several figures can illustrate the status and development prospects of antibody drugs: In 2010, the total sales of monoclonal antibody drugs for global treatment reached 44 billion US dollars. Monoclonal antibody diagnostic and research reagents worth US$10 billion, the total market for monoclonal antibody drugs has reached US$55 billion, and the total market for monoclonal antibody drugs in 2011 has reached US$62.8 billion. In 2009 and 2008, the figures were US$40 billion and US$37 billion respectively (data fro...

Claims

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Application Information

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IPC IPC(8): C07K16/18C07K1/22C07K1/18
Inventor 朱健万艳孙旭东陈广勇鲁钱达瞿丽莉谢晋胡可可高金湖沈克强周伟昌陈智胜
Owner 上海药明生物医药有限公司
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