Expression, purification, and characterization of rhTyrRS
Yongjiang Lang, Yanling Zhang, Ling Zhan
1The Key Laboratory of Molecular Medicine, Ministry of Education, Fudan University, Shanghai, P,R, China. minyu@shmu.edu.cn.
BMC Biotechnology
|July 17, 2014
Summary
A new industrial production method yields high-purity recombinant human tyrosyl-tRNA synthetase (rhTyrRS) for research. This engineered protein, rhTyrRS, demonstrates cytokine activity, paving the way for preclinical studies.
Area of Science:
- Biochemistry
- Molecular Biology
- Protein Engineering
Background:
- Aminoacyl-tRNA synthetases (AARSs) are crucial for protein synthesis.
- AARSs have emerging roles in signal transduction, influencing thrombopoiesis and inflammation.
- Engineered recombinant human tyrosyl-tRNA synthetase (rhTyrRS) exhibits unmasked cytokine activity.
Purpose of the Study:
- To develop an industrial production method for high-yield, high-purity, quality, and potency rhTyrRS.
- To enable preclinical research using rhTyrRS.
Main Methods:
- Codon-optimized rhTyrRS was expressed in Escherichia coli under fermentation.
- A three-step purification process involving cation exchange, gel filtration, and anion exchange chromatography was employed.
- Biological activity was assessed via aminoacylation and IL-8 release in HL-60 cells.
Main Results:
- High-purity, soluble rhTyrRS was successfully produced.
- A robust purification strategy was established.
- Methods for assessing rhTyrRS biological activity were developed.
Conclusions:
- The characterized rhTyrRS is suitable for pharmacodynamic and pharmacokinetic studies.
- The developed production method ensures protein quality for research applications.
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