Abstract
It has been a conventional notion that cytoplasmic recombinant expression leads to either soluble protein or inclusion bodies. In the latter case, it was always assumed that proteins in inclusion bodies (IBs) are more or less unfolded and hence require complete denaturing condition for solubilization, which uses strong detergents, urea or guanidine hydrochloride. However, we often observe distribution of expressed proteins in both soluble and insoluble fractions. In such expression, IBs are often loose and of flocculate morphology. We believe that such distribution is due to association of near native structures of the expressed proteins, which cause either aggregation into insoluble fractions or unstable soluble proteins. In our experience, although not reported by others, interleukin-1α, interferon-γ, tumor necrosis factors, fibroblast growth factors, His-tagged fyn kinase and many other proteins showed such behavior. If this occurs, we have experienced problems of instability, low yield and insolubility whether purification is done from the soluble fraction or by refolding of IBs. Arginine has shown great promise in non-denaturaing solubilization of some of these proteins we have tested.
Keywords: Non-denaturing solubilization, inclusion bodies, arginine, GFP, FGF-20
Current Pharmaceutical Biotechnology
Title: Non-Denaturing Solubilization of Inclusion Bodies
Volume: 11 Issue: 3
Author(s): Kouhei Tsumoto, Ryota Abe, Daisuke Ejima and Tsutomu Arakawa
Affiliation:
Keywords: Non-denaturing solubilization, inclusion bodies, arginine, GFP, FGF-20
Abstract: It has been a conventional notion that cytoplasmic recombinant expression leads to either soluble protein or inclusion bodies. In the latter case, it was always assumed that proteins in inclusion bodies (IBs) are more or less unfolded and hence require complete denaturing condition for solubilization, which uses strong detergents, urea or guanidine hydrochloride. However, we often observe distribution of expressed proteins in both soluble and insoluble fractions. In such expression, IBs are often loose and of flocculate morphology. We believe that such distribution is due to association of near native structures of the expressed proteins, which cause either aggregation into insoluble fractions or unstable soluble proteins. In our experience, although not reported by others, interleukin-1α, interferon-γ, tumor necrosis factors, fibroblast growth factors, His-tagged fyn kinase and many other proteins showed such behavior. If this occurs, we have experienced problems of instability, low yield and insolubility whether purification is done from the soluble fraction or by refolding of IBs. Arginine has shown great promise in non-denaturaing solubilization of some of these proteins we have tested.
Export Options
About this article
Cite this article as:
Tsumoto Kouhei, Abe Ryota, Ejima Daisuke and Arakawa Tsutomu, Non-Denaturing Solubilization of Inclusion Bodies, Current Pharmaceutical Biotechnology 2010; 11 (3) . https://dx.doi.org/10.2174/138920110791111924
DOI https://dx.doi.org/10.2174/138920110791111924 |
Print ISSN 1389-2010 |
Publisher Name Bentham Science Publisher |
Online ISSN 1873-4316 |
Call for Papers in Thematic Issues
Artificial Intelligence in Bioinformatics
Bioinformatics is an interdisciplinary field that analyzes and explores biological data. This field combines biology and information system. Artificial Intelligence (AI) has attracted great attention as it tries to replicate human intelligence. It has become common technology for analyzing and solving complex data and problems and encompasses sub-fields of machine ...read more
Latest Advancements in Biotherapeutics
The scope of this thematic issue is to comprehensively explore the rapidly evolving landscape of biotherapeutics, emphasizing breakthroughs in precision medicine. Encompassing diverse therapeutic modalities, the issue will delve into the latest developments in monoclonal antibodies, CRISPR/Cas gene editing, CAR-T cell therapies, and innovative drug delivery systems, such as nanoparticle-based ...read more
- 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
-
Radioprotective Gene Therapy
Current Gene Therapy Neuropathic Pain: Models and Mechanisms
Current Pharmaceutical Design Opinion Paper: Promise and Pragmatism in Clinical Microbiome Research
Mini-Reviews in Medicinal Chemistry Emerging In Vitro Tools to Evaluate Cytochrome P450 and Transporter-Mediated Drug-Drug Interactions
Current Drug Discovery Technologies Ethnicity and Inflammatory Pathways - Implications for Vascular Disease, Vascular Risk and Therapeutic Intervention
Current Medicinal Chemistry The Applicability of mTOR Inhibition in Solid Tumors
Current Cancer Drug Targets Therapeutic Hypothermia and Neuroprotection in Acute Neurological Disease
Current Medicinal Chemistry Editorial: Computer-Assisted Approaches as Decision Support Systems in the Overall Strategy of Combating Emerging Diseases: Some Comments Regarding Drug Design, Vaccinomics, and Genomic Surveillance of the Zika Virus
Current Computer-Aided Drug Design Cardiovascular Risk Factors in Chronic Inflammatory Rheumatic Diseases: Modern Assessment and Diagnosis
Current Vascular Pharmacology A Mixed Anti-Inflammatory and Pro-Inflammatory Response Associated with a High Dose of Corticosteroids
Current Molecular Medicine Deciphering G Protein-Coupled Receptor Biology with Fluorescence-based Methods
Current Pharmaceutical Biotechnology Vitamin, Mineral, and Drug Absorption Following Bariatric Surgery
Current Drug Metabolism Synthesis of Some New Hydroxytriazenes and their Antimicrobial and Anti-inflammatory Activities
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Editorial (Thematic Issue: The Changing Face of Metabolic Syndrome and its Components in the Light of Current Knowledge)
Current Vascular Pharmacology Antibiotics in Paediatrics: A Boon or a Bane?
Anti-Infective Agents Retroviral Protein Transfer: Falling Apart to Make an Impact
Current Gene Therapy Vitamin D/VDR in Acute Kidney Injury: A Potential Therapeutic Target
Current Medicinal Chemistry Advances in Vaccine Adjuvants For Infectious Diseases
Current HIV Research Astrocytes: Implications for Neuroinflammatory Pathogenesis of Alzheimers Disease
Current Alzheimer Research Subject Index To Volume 5
CNS & Neurological Disorders - Drug Targets