Related Experiment Video
Updated: Jul 4, 2026
07:09
A Convenient and General Expression Platform for the Production of Secreted Proteins from Human Cells
Published on: July 31, 2012
High efficient mammalian expression and secretion of a functional humanized single-chain Fv/human interleukin-2
Yue-Chun Shen1, Xue-Hao Wang, Xiao-Ming Wang
1Division of Cardiology, Department of Internal Medicine, The University of Michigan Medical School, Ann Arbor, 48109, USA.
World Journal of Gastroenterology
|June 29, 2006
Summary
Researchers engineered a bispecific antibody-Interleukin-2 (IL-2) fusion protein in mammalian cells. This novel fusion protein effectively targets HER-2/neu positive tumors, retaining IL-2
Area of Science:
- Biotechnology and Genetic Engineering
- Immunology and Cancer Therapy
Background:
- Development of targeted cancer therapies is crucial for improving treatment efficacy.
- Bispecific fusion proteins offer a strategy to combine therapeutic payloads with tumor-targeting moieties.
- Interleukin-2 (IL-2) has immunostimulatory properties but systemic administration can cause toxicity.
Purpose of the Study:
- To construct and produce a recombinant bispecific humanized single-chain variable fragment (sFv) /Interleukin-2 (IL-2) fusion protein.
- To evaluate the efficiency of mammalian cells in producing and secreting active sFv/IL-2.
- To assess the therapeutic potential of targeting IL-2 to HER-2/neu positive tumors.
Main Methods:
- Genetic engineering of the sFv/IL-2 fusion protein.
- Transfection into mammalian cells (293 cell line) for protein production.
- Assays to measure IL-2 activity (cell proliferation, cytotoxicity) and antigen-binding specificity (ELISA).
Main Results:
- High-efficiency expression of the sFv/IL-2 fusion protein achieved (up to 102 +/- 4.2 mg/L).
- The fusion protein demonstrated retained IL-2 immunostimulatory activities.
- Antigen-binding specificity against HER-2/neu (p185) was confirmed.
Conclusions:
- Successful large-scale preparation of the recombinant humanized sFv antibody/IL-2 fusion protein using 293 cells.
- This fusion protein enables targeted delivery of IL-2 to p185-positive tumors.
- Potential for reduced systemic toxicity and immunogenicity compared to conventional IL-2 therapy.
