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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