Abstract
An efficient preparation of Periplaneta americana nymphae allergen, Cr PI (54 kDa) is described. It was expressed as a GST-tag fusion protein in Escherichia coli, strain BL21 (DE3). Expression of recombinant Cr PI (rCr PI), denaturation/ renaturation of the inclusion bodies and the effects of protein and L-arginine concentration on inclusion body aggregation were optimized. The fusion protein was purified by affinity chromatography and size exclusion chromatography, and Cr PI fusion protein was purified to > 95%. rCr PI bound strongly to IgE in the sera of individuals with cockroach allergies as shown by western blot and ELISA. Highly refolded and purified recombinant protein was obtained, providing a basis for the large-scale preparation of Cr PI allergen.
Keywords: Periplaneta americana, Cr PI allergen, high-level expression, inclusion bodies, immunological characterization
Protein & Peptide Letters
Title: Expression, Purification, and Immunological Characterization of Cr PI
Volume: 14 Issue: 9
Author(s): Hai-Qiang Wu, Zhi-Gang Liu, Pi-Xin Ran, Zhen-Wen Zhou and Bo Gao
Affiliation:
Keywords: Periplaneta americana, Cr PI allergen, high-level expression, inclusion bodies, immunological characterization
Abstract: An efficient preparation of Periplaneta americana nymphae allergen, Cr PI (54 kDa) is described. It was expressed as a GST-tag fusion protein in Escherichia coli, strain BL21 (DE3). Expression of recombinant Cr PI (rCr PI), denaturation/ renaturation of the inclusion bodies and the effects of protein and L-arginine concentration on inclusion body aggregation were optimized. The fusion protein was purified by affinity chromatography and size exclusion chromatography, and Cr PI fusion protein was purified to > 95%. rCr PI bound strongly to IgE in the sera of individuals with cockroach allergies as shown by western blot and ELISA. Highly refolded and purified recombinant protein was obtained, providing a basis for the large-scale preparation of Cr PI allergen.
Export Options
About this article
Cite this article as:
Wu Hai-Qiang, Liu Zhi-Gang, Ran Pi-Xin, Zhou Zhen-Wen and Gao Bo, Expression, Purification, and Immunological Characterization of Cr PI, Protein & Peptide Letters 2007; 14 (9) . https://dx.doi.org/10.2174/092986607782110266
DOI https://dx.doi.org/10.2174/092986607782110266 |
Print ISSN 0929-8665 |
Publisher Name Bentham Science Publisher |
Online ISSN 1875-5305 |

- Author Guidelines
- Bentham Author Support Services (BASS)
- 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
-
Personalised Genetic Intervention for Duchenne Muscular Dystrophy: Antisense Oligomers and Exon Skipping
Current Molecular Pharmacology Application of Human Placental Villous Tissue Explants to Study ABC Transporter Mediated Efflux of 2,4-Dinitrophenyl-S-Glutathione
Current Pharmaceutical Biotechnology Merging Traditional Chinese Medicine with Modern Drug Discovery Technologies to Find Novel Drugs and Functional Foods
Current Drug Discovery Technologies Sleep in Pediatric Pulmonary Diseases
Current Respiratory Medicine Reviews Inhaled Corticosteroids In Treatment of Cystic Fibrosis
Anti-Inflammatory & Anti-Allergy Agents in Medicinal Chemistry Liposomes as Nanovaccine Delivery Systems
Current Topics in Medicinal Chemistry Paracetamol-Induced Stevens Johnson Syndrome and Cholestatic Hepatitis
Current Drug Safety Supercritical Fluid Chromatographic Method for Montelukast: Application in Content Uniformity and Degradation Study
Current Analytical Chemistry Peripheral TRPV1 Receptors As Targets for Drug Development: New Molecules and Mechanisms
Current Pharmaceutical Design Relevance of the Deletion Polymorphisms of the Glutathione S-Transferases GSTT1 and GSTM1 in Pharmacology and Toxicology
Current Drug Metabolism Understanding the Multifaceted Role of Ectonucleotide Pyrophosphatase/Phosphodiesterase 2 (ENPP2) and its Altered Behaviour in Human Diseases
Current Molecular Medicine Controlled Parenteral Formulations: An Efficacious and Favourable Way to Deliver the Anti-psychotic Drugs
Current Psychiatry Research and Reviews Biodiversity and Chemodiversity: Future Perspectives in Bioprospecting
Current Drug Targets Molecular Recognition of Tachykinin Receptor Selective Agonists: Insights from Structural Studies
Mini-Reviews in Medicinal Chemistry Editorial:
Drug Metabolism Letters Potential Genetic Therapies for Acute Lung Injury
Current Gene Therapy Metabotropic Purinergic Receptors in Lipid Membrane Microdomains
Current Medicinal Chemistry Editorial from Guest Editors [Hot Topic: Viral Bronchiolitis: The Year in Review (Guest Editors: Louis Bont and Jan Kimpen)]
Current Respiratory Medicine Reviews Determining Effective Diabetic Care; A Multicentre - Longitudinal Interventional Study
Current Pharmaceutical Design Regulation of the Endoplasmic Reticulum Ca2+-Store in Cancer
Anti-Cancer Agents in Medicinal Chemistry