On-column refolding purification of DT389-hIL13 recombinant protein expressed in Escherichia coli.
Summary of "On-column refolding purification of DT389-hIL13 recombinant protein expressed in Escherichia coli."
Protein refolding is a bottleneck in the production of therapeutic proteins from inclusion bodies. In recent years, several studies have described on-column refolding of recombinant proteins. DT389-hIL13 is a recombinant protein that targets the glioma. In our study, the recombinant protein DT389-hIL13 was expressed in Escherichia coli (E. coli). The isolated inclusion bodies were refolded using size exclusion chromatography (SEC) and further purified using anion exchange chromatography. Three different methods of SEC on-column refolding were studied. In vitro tests on U251 cells showed that the recombinant protein could effectively inhibit the proliferation of U251 cells, especially the protein refolded by urea and pH gradient method. The half-maximal inhibitory concentration (IC50) of 0.887 nM was achieved with this new method, unlike an IC50 of 11.4 nM achieved in the non-gradient method.
Affiliation
College of Life Sciences and Bioengineering, Beijing Jiaotong University, Beijing 100044, China.
Journal Details
This article was published in the following journal.
Name: Protein expression and purification
ISSN: 1096-0279
Pages:
Links
- PubMed Source: http://www.ncbi.nlm.nih.gov/pubmed/20851185
- DOI: http://dx.doi.org/10.1016/j.pep.2010.09.009
Medical and Biotech [MESH] Definitions
Protein Refolding
Conformational transitions of a protein from unfolded states to a more folded state.
Artificial Gene Fusion
The in vitro fusion of GENES by RECOMBINANT DNA techniques to analyze protein behavior or GENE EXPRESSION REGULATION, or to merge protein functions for specific medical or industrial uses.
Kangai-1 Protein
A widely expressed transmembrane glycoprotein that functions as a METASTASIS suppressor protein. It is under-expressed in a variety of human NEOPLASMS.
Amyloid
A type of extracellularly deposited substance composed of an amyloid protein and additional components including HEPARAN SULFATE PROTEOGLYCAN; LAMININ; COLLAGEN TYPE IV; SERUM AMYLOID P-COMPONENT; and APOLIPOPROTEINS E which together form characteristic amyloid fibrils. The core of amyloid fibrils is formed by the stacking of overlapping beta-pleated sheet domains of the amyloid protein. There are many different amyloid proteins that have been found forming the core of the fibrils in vivo. However, amyloid can be formed from any protein that exposes beta-pleated strand conformations during unfolding or refolding. A common characteristic of amyloid is the ability to bind such dyes as CONGO RED and thioflavine.
Retinoblastoma-binding Protein 1
A ubiquitously expressed regulatory protein that contains a retinoblastoma protein binding domain and an AT-rich interactive domain. The protein may play a role in recruiting HISTONE DEACETYLASES to the site of RETINOBLASTOMA PROTEIN-containing transcriptional repressor complexes.
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