title: Purification of recombinant ovalbumin from inclusion bodies of Escherichia coli.
abstract: Recombinant ovalbumin expressed in bacterial host is essentially free from post-translational modifications and can be useful in understanding the structure-function relationship of the protein. In this study, ovalbumin was expressed in Escherichia coli in the form of inclusion bodies. Ovalbumin inclusion bodies were solubilized using urea and refolded by decreasing the urea concentration by dilution. Refolded protein was purified by anion exchange chromatography. Overall recovery of purified recombinant ovalbumin from inclusion bodies was about 30% with 98% purity. Purified recombinant ovalbumin was characterized by mass spectrometry, circular dichroism and fluorescence spectroscopy. Recombinant ovalbumin was shown to be resistant to trypsin using protease resistance assay. This indicated proper refolding of ovalbumin from inclusion bodies of E. coli. This method provides a simple way of producing ovalbumin free of post-translational modifications.
authors.affiliation: Product Development Cell, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Vaibhav
authors.initials: V
authors.lastname: Upadhyay
authors.affiliation: Product Development Cell, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India.
authors.forename: Anupam
authors.initials: A
authors.lastname: Singh
authors.affiliation: Product Development Cell, National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110067, India. Electronic address: amulya@nii.res.in.
authors.forename: Amulya K
authors.initials: AK
authors.lastname: Panda
mesh_terms: D004926:Escherichia coli / Q000737:chemistry*; D002479:Inclusion Bodies / Q000737:chemistry*; D010047:Ovalbumin / Q000737:chemistry / Q000302:isolation & purification; D058849:Protein Refolding; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification
chemical_list: D011994:Recombinant Proteins; D010047:Ovalbumin
keywords: Inclusion bodies; Ovalbumin; Refolding and purification; Solubilization