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Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘

Title
Poly-lysine이 연결된 hEGF와 angiogenin의 융합단백질의 고체상 재접힘
Other Titles
Solid-Phase Refolding of Poly-Lysine Tagged Fusion Protein of hEGF and Angiogenin
Author
이은규
Keywords
refolding; solid-phase refolding; inclusion body; fusion protein; heparin; cationic tagging
Issue Date
2002-04
Publisher
한국생물공학회
Citation
KSBB Journal, v. 17, no. 2, page. 153-157
Abstract
A fusion protein, consisting of a human epidermal growth factor 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 an inclusion body in recombinant E. coli, yet when the conventional solution-phase refolding process was used the refolding yield was very low due to severe aggregation, probably because of the opposite surface charge resulting from the vastly different pI values of each domain. Accordingly the solid-phase refolding process, which exploits the ionic interactions between a solid matrix and the protein, was tried, however the ionic binding yield was also very low regardless of the resins and pH conditions used. Therefore, to provide a higher affinity toward the solid matrix, six lysine residues were tagged to the N-terminus of the hEGF domain. When cation exchange resins, such as heparin- or CM-Sepharose, were used as the matrix, the adsorption capacity increased 2.5 ~3-fold and the subsequent refolding yield increased nearly 15-fold compared to the conventional process. A similar result was also obtained when an Ni-NTA metal affinity resin was used.
URI
https://www.earticle.net/Article/A101117https://repository.hanyang.ac.kr/handle/20.500.11754/157242
ISSN
1225-7117
Appears in Collections:
COLLEGE OF ENGINEERING SCIENCES[E](공학대학) > BIONANO ENGINEERING(생명나노공학과) > Articles
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