112 80

Solid-phase Refolding of Poly-lysine Tagged Fusion Protein of hEGF and Angiogenin

Title
Solid-phase Refolding of Poly-lysine Tagged Fusion Protein of hEGF and Angiogenin
Author
이은규
Keywords
refolding; solid-phase refolding; inclusion body; fusion protein; cationic tagging; heparin
Issue Date
2002-01
Publisher
한국생물공학회
Citation
Biotechnology and Bioprocess Engineering, v. 7, issue. 1, page. 1-5
Abstract
A fusion protein, consisting of a human epidermal growth factor (hEGF) as the recognition domain and human angiogenin as the toxin domain, can be used as a targeted therapeutic against breast cancer cells among others. The fusion protein was expressed as inclusion body in recombinantE. coli, and when the conventional, solution-phase refolding process was used the refolding yield was very low due to severe aggregation. It was probably because of the opposite electric charge at a neutral pH resulting from the vastly different pI values of each domain. The solidphase refolding process that exploited the ionic interactions between ionic exchanger surface and the fusion protein was tried, but the adsorption yield was also very low, below 30%, regardless of the resins and pH conditions used. Therefore, to provide a higher ionic affinity toward the solid matrix, six lysine residues were tagged to theN-terminus of the hEGF domain. When heparin-Sepharose was used as the matrix, the adsorption capacity increased 2.5–3 times to about 88%. Besides the intrinsic affinity of angiogenin to heparin the poly-lysine tag provided additional ionic affinity. And the subsequent refolding yield increased nearly 13-fold, fromc.a 4.8% in the conventional refolding of the untagged fusion protein to 63.6%. The process was highly reproducible. The refolded protein in the column eluate retained R Nase bioactivity, of angiogenin.
URI
https://link.springer.com/article/10.1007/BF02935871https://repository.hanyang.ac.kr/handle/20.500.11754/156889
ISSN
1226-8372
DOI
10.1007/BF02935871
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
Files in This Item:
2002.01-이은규-Solid-phase Refolding of Poly-lysine Tagged Fusion Protein of hEGF and Angiogenin.pdfDownload
Export
RIS (EndNote)
XLS (Excel)
XML


qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE