title: Soni-removal of nucleic acids from inclusion bodies.
abstract: Inclusion bodies (IBs) are commonly formed in Escherichia coli due to over expression of recombinant proteins in non-native state. Isolation, denaturation and refolding of these IBs is generally performed to obtain functional protein. However, during this process IBs tend to form non-specific interactions with sheared nucleic acids from the genome, thus getting carried over into downstream processes. This may hinder the refolding of IBs into their native state. To circumvent this, we demonstrate a methodology termed soni-removal which involves disruption of nucleic acid-inclusion body interaction using sonication; followed by solvent based separation. As opposed to conventional techniques that use enzymes and column-based separations, soni-removal is a cost effective alternative for complete elimination of buried and/or strongly bound short nucleic acid contaminants from IBs.
authors.affiliation: Institute of Molecular Medicine (IMM), The University of Texas Health Sciences Centre at Houston, 1825 Pressler Street, Houston, TX 77030, USA; Protein Technology Core (PTC), Centre for Cellular and Molecular Platforms (C-CAMP), NCBS-TIFR, GKVK Post, Bellary Road, Bangalore 560065, India. Electronic address: munish@ccamp.res.in.
authors.forename: Muniasamy
authors.initials: M
authors.lastname: Neerathilingam
authors.affiliation: Protein Technology Core (PTC), Centre for Cellular and Molecular Platforms (C-CAMP), NCBS-TIFR, GKVK Post, Bellary Road, Bangalore 560065, India.
authors.forename: Sumukh
authors.initials: S
authors.lastname: Mysore
authors.affiliation: Institute of Molecular Medicine (IMM), The University of Texas Health Sciences Centre at Houston, 1825 Pressler Street, Houston, TX 77030, USA.
authors.forename: Sai Hari A
authors.initials: SH
authors.lastname: Gandham
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D003716:Dengue Virus / Q000737:chemistry; D018960:Hyaluronan Receptors / Q000302:isolation & purification; D002479:Inclusion Bodies / Q000378:metabolism*; D009696:Nucleic Acids / Q000302:isolation & purification* / Q000378:metabolism; D011489:Protein Denaturation; D017510:Protein Folding; D011994:Recombinant Proteins / Q000302:isolation & purification; D012995:Solubility; D013010:Sonication; D014759:Viral Envelope Proteins / Q000302:isolation & purification
chemical_list: D018960:Hyaluronan Receptors; D009696:Nucleic Acids; D011994:Recombinant Proteins; D014759:Viral Envelope Proteins
keywords: Economic; Inclusion bodies purification; Nucleic acid contamination; Soni-Removal; Sonication