Cost-effective production of recombinant human interleukin 24 by lactose induction and a two-step denaturing and one-step refolding method

Recombinant human interleukin 24 (rhIL24) is a member of the interleukin 10 (IL10) family of cytokines with novel therapeutic properties. Human IL24 possesses three N glycosylation sites and a disulfide bridge. The cost and composition of culture media is critical for commercial-scale production of...

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Published in:Journal of industrial microbiology & biotechnology 2014, Vol.41 (1), p.135-142
Main Authors: Amirzada, Muhammad Imran, Yu, Minglei, Gong, Xiaohai, Chen, Yun, Zhu, Ruiyu, Lei, Jianyong, Jin, Jian
Format: Article
Language:English
Subjects:
Analysis
Biochemistry
Bioinformatics
Biological activity
Biological and medical sciences
Biomedical and Life Sciences
Biosynthesis
Biotechnology
Cell culture
Cell Culture and Bioengineering
Cell Line, Tumor
Chloride
Culture media
Cytokines
disulfide bonds
E coli
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Fermentation
Fundamental and applied biological sciences. Psychology
Genetic Engineering
Genetic recombination
Glucose
Glycerol
hepatoma
Humans
Inorganic Chemistry
interleukin-10
Interleukins - biosynthesis
Interleukins - genetics
Interleukins - pharmacology
Lactose
Lactose - metabolism
Life Sciences
Liver cancer
medicinal properties
Microbiology
Potassium
product quality
Protein Denaturation
Protein folding
Protein Refolding
Proteins
recombinant proteins
Recombinant Proteins - biosynthesis
Recombinant Proteins - chemistry
Recombinant Proteins - economics
Recombinant Proteins - pharmacology
Sodium
Studies
Tandem Mass Spectrometry
Toxicity
Yeast
yeast extract
Yeasts
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cited_by cdi_FETCH-LOGICAL-c539t-49f10118fcce769bde132278317ab6d9386e8edae7448ad98f59d61468f4eaf23
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container_end_page 142
container_issue 1
container_start_page 135
container_title Journal of industrial microbiology & biotechnology
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creator Amirzada, Muhammad Imran
Yu, Minglei
Gong, Xiaohai
Chen, Yun
Zhu, Ruiyu
Lei, Jianyong
Jin, Jian
description Recombinant human interleukin 24 (rhIL24) is a member of the interleukin 10 (IL10) family of cytokines with novel therapeutic properties. Human IL24 possesses three N glycosylation sites and a disulfide bridge. The cost and composition of culture media is critical for commercial-scale production of recombinant proteins in E. coli. Addition of yeast extract and glucose to medium enhances rhIL24 production, and the use of lactose instead of IPTG for induction drops the cost and decreases toxicity. In addition, a two-step denaturing and one-step refolding (2DR) strategy improves rhIL24 production. The 2DR strategy replaces a more conventional approach for protein solubilization and refolding. LC–MS/MS provides definitive identification and quantitative information on rhIL24. Single-step purified rhIL24 displayed biological activity on HepG2 hepatocellular carcinoma cells, but no effect on L02 cells. Proliferation analysis suggests that rhIL24 may have potential use as a medication. In the present study, we developed a simple process for producing quality product with high purity. The expression and purification of rhIL24 described here may be a step towards inexpensive large-scale production.
doi_str_mv 10.1007/s10295-013-1367-2
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Human IL24 possesses three N glycosylation sites and a disulfide bridge. The cost and composition of culture media is critical for commercial-scale production of recombinant proteins in E. coli. Addition of yeast extract and glucose to medium enhances rhIL24 production, and the use of lactose instead of IPTG for induction drops the cost and decreases toxicity. In addition, a two-step denaturing and one-step refolding (2DR) strategy improves rhIL24 production. The 2DR strategy replaces a more conventional approach for protein solubilization and refolding. LC–MS/MS provides definitive identification and quantitative information on rhIL24. Single-step purified rhIL24 displayed biological activity on HepG2 hepatocellular carcinoma cells, but no effect on L02 cells. Proliferation analysis suggests that rhIL24 may have potential use as a medication. In the present study, we developed a simple process for producing quality product with high purity. 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subjects Analysis
Biochemistry
Bioinformatics
Biological activity
Biological and medical sciences
Biomedical and Life Sciences
Biosynthesis
Biotechnology
Cell culture
Cell Culture and Bioengineering
Cell Line, Tumor
Chloride
Culture media
Cytokines
disulfide bonds
E coli
Escherichia coli
Escherichia coli - genetics
Escherichia coli - metabolism
Fermentation
Fundamental and applied biological sciences. Psychology
Genetic Engineering
Genetic recombination
Glucose
Glycerol
hepatoma
Humans
Inorganic Chemistry
interleukin-10
Interleukins - biosynthesis
Interleukins - genetics
Interleukins - pharmacology
Lactose
Lactose - metabolism
Life Sciences
Liver cancer
medicinal properties
Microbiology
Potassium
product quality
Protein Denaturation
Protein folding
Protein Refolding
Proteins
recombinant proteins
Recombinant Proteins - biosynthesis
Recombinant Proteins - chemistry
Recombinant Proteins - economics
Recombinant Proteins - pharmacology
Sodium
Studies
Tandem Mass Spectrometry
Toxicity
Yeast
yeast extract
Yeasts
title Cost-effective production of recombinant human interleukin 24 by lactose induction and a two-step denaturing and one-step refolding method
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