Separation and purification method of recombinant human interferon beta 1a

Abstract

The invention discloses a separation and purification method of recombinant human interferon beta 1a. The separation and purification method comprises the following steps: 1) carrying out blue gel chromatography on a sample, collecting active peaks of the recombinant human interferon beta 1a; 2) carrying out ultrafiltration and desalination on a product obtained by the blue gel chromatography; 3)carrying out Q-Sepharose Fast Flow anion exchange chromatography on the ultrafiltration and desalination product, balancing a column by using 20mmol / / L Tris-HCl buffer liquid with the pH of 8.3-8.7, after sampling, eluting with the buffer liquid to a base line, and then eluting by using 20mmol / L NaCl-containing Tris-HCl buffer liquid with the pH of 8.3-8.7, and collecting the active peaks of the recombinant human interferon beta 1a; and 4) carrying out S-200 molecular sieve chromatography on the product obtained by the anion exchange chromatography so as to obtain the recombinant human interferon beta 1a. By using the purification method, a target protein has small possibility of inactivation, and the purity and biology specific activity of the target protein obtained by purification are high.

Description

technical field [0001] The invention relates to the field of purification of genetically engineered recombinant proteins, in particular to a method for separating and purifying recombinant human interferon beta 1a by using anion exchange chromatography technology. Background technique [0002] Human interferon β (IFN-β) is an important cytokine with anti-virus, anti-tumor, and immune regulation effects, mainly produced by fibroblasts and some epithelial cells, common interferon-inducing agents, such as viruses, double-stranded Both RNA and some microorganisms can induce the production of IFN-β. Natural IFN-β is a glycoprotein (sugar content accounts for about 20%), with a relative molecular mass of about 20,000, consisting of 166 amino acids, and a signal peptide consisting of 21 amino acids, cut between S-M. The 17th, 31st, and 141st positions of the molecule are Cys, and the disulfide bond formed between the 31st and 141st positions is necessary for its biological activit...

AI Technical Summary

Problems solved by technology

[0004] The currently reported purification method uses CM cation chromatography, the pH of the buffer solution is changed between 7.2 and 5.0, the protein is easily deformed, the activity loss is large, the operation is cumbersome, and the process is difficult to scale up

Images