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Soni-removal of nucleic acids from inclusion bodies. [Protein Technology Core]

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TitleSoni-removal of nucleic acids from inclusion bodies. [Protein Technology Core]
Publication TypeJournal Article
Year of Publication2014
AuthorsNeerathilingam M, Mysore S, Gandham SHari A
JournalBiochem Biophys Res Commun
Volume448
Issue1
Pagination45-9
Date Published2014 May 23
ISSN1090-2104
KeywordsAntigens, CD44, Cell Fractionation, Dengue Virus, Inclusion Bodies, Nucleic Acids, Protein Denaturation, Protein Folding, Recombinant Proteins, Solubility, Sonication, Viral Envelope Proteins
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.

DOI10.1016/j.bbrc.2014.04.049
Alternate JournalBiochem. Biophys. Res. Commun.
PubMed ID24747565