Abstract
In the current study, a mutant recombinant human interleukin-2 (MhIL-2) was generated using site-directed mutagenesis. The bacteria transformed with plasmid pET15b-MhIL-2 were cultured in LB medium containing 0.6mM IPTG for 8 hours at 27°C. Approximately 90% of His-MhIL-2 was efficiently expressed in soluble form. Purification efficiency was optimized using a number of strategies, including nickel ion chelating chromatography, desalting chromatography, thrombin cleavage and Superdex 75 gel filtration chromatography. The final product had > 95% purity. PBMCs, CD4+ and CD8+ T cell proliferation assays revealed that one such mutant has identical functional property to the wild-type hIL-2. In summary, we generated a mutant hIL-2 that is functionally identical to wild-type hIL-2.
Keywords: Biological activity, chromatography, expression, interleukin-2, mutant, purification, soluble form
Protein & Peptide Letters
Title: Expression, Purification, and Characterization of a Functional Mutant Recombinant Human Interleukin-2
Volume: 17 Issue: 10
Author(s): Mingjun Liu, Bin Wang, Guirong Sun, Dongmeng Qian, Zhiyong Yan, Xuxia Song and Shouyi Ding
Affiliation:
Keywords: Biological activity, chromatography, expression, interleukin-2, mutant, purification, soluble form
Abstract: In the current study, a mutant recombinant human interleukin-2 (MhIL-2) was generated using site-directed mutagenesis. The bacteria transformed with plasmid pET15b-MhIL-2 were cultured in LB medium containing 0.6mM IPTG for 8 hours at 27°C. Approximately 90% of His-MhIL-2 was efficiently expressed in soluble form. Purification efficiency was optimized using a number of strategies, including nickel ion chelating chromatography, desalting chromatography, thrombin cleavage and Superdex 75 gel filtration chromatography. The final product had > 95% purity. PBMCs, CD4+ and CD8+ T cell proliferation assays revealed that one such mutant has identical functional property to the wild-type hIL-2. In summary, we generated a mutant hIL-2 that is functionally identical to wild-type hIL-2.
Export Options
About this article
Cite this article as:
Liu Mingjun, Wang Bin, Sun Guirong, Qian Dongmeng, Yan Zhiyong, Song Xuxia and Ding Shouyi, Expression, Purification, and Characterization of a Functional Mutant Recombinant Human Interleukin-2, Protein & Peptide Letters 2010; 17 (10) . https://dx.doi.org/10.2174/092986610792231474
DOI https://dx.doi.org/10.2174/092986610792231474 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |
- Author Guidelines
- Graphical Abstracts
- Fabricating and Stating False Information
- Research Misconduct
- Post Publication Discussions and Corrections
- Publishing Ethics and Rectitude
- Increase Visibility of Your Article
- Archiving Policies
- Peer Review Workflow
- Order Your Article Before Print
- Promote Your Article
- Manuscript Transfer Facility
- Editorial Policies
- Allegations from Whistleblowers
Related Articles
-
Patent Selections
Recent Patents on Inflammation & Allergy Drug Discovery Colchicine: An Old Wine in a New Bottle?
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Dietary Intake and Bone Status with Aging
Current Pharmaceutical Design α7 Nicotinic Acetylcholine Receptor Subunit in Angiogenesis and Epithelial to Mesenchymal Transition
Current Drug Targets Allergen-Induced Inflammation
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Possible Targets of Herbals for Type 3 Diabetes: A Review
Current Traditional Medicine Subject Index to Volume 2
Current Medicinal Chemistry - Cardiovascular & Hematological Agents Diagnosis, Prevention and Treatment of Aspirin-Induced Asthma and Rhinitis
Current Drug Targets - Inflammation & Allergy Clinical and Forensic Signs Related to Cocaine Abuse
Current Drug Abuse Reviews Long-Term Multimodal Therapy (Verapamil Associated with Propolis, Blueberry, Vitamin E and Local Diclofenac) on Patients with Peyronie's Disease (Chronic Inflammation of the Tunica Albuginea). Results of a Controlled Study
Inflammation & Allergy - Drug Targets (Discontinued) Subject index to volume 1
Current HIV Research Small Non-Coding RNAs as Novel Therapeutics
Current Molecular Medicine High-Level Secretory Expression and Purification of Recombinant Human Interleukin 1 Beta in Pichia pastoris
Protein & Peptide Letters Autoimmune Diseases and Infections: A Controversial Relationship
Current Immunology Reviews (Discontinued) Role of the Hypoxic Microenvironment in the Antitumor Activity of Tyrosine Kinase Inhibitors
Current Medicinal Chemistry Molecular Adjuvants Based on Plasmids Encoding Protein Aggregation Domains Affect Bone Marrow Niche Homeostasis
Current Gene Therapy Genomic Instability in Cancer: Molecular Mechanisms and Therapeutic Potentials
Current Pharmaceutical Design One-Compound-Multi-Target: Combination Prospect of Natural Compounds with Thrombolytic Therapy in Acute Ischemic Stroke
Current Neuropharmacology Pharmacology of Rhein and Advancement in the Synthesis of Its Derivatives
Current Traditional Medicine Whole Genome Re-sequencing of Soybean Accession EC241780 Providing Genomic Landscape of Candidate Genes Involved in Rust Resistance
Current Genomics