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Eliminating the six N ‐terminal amino acids of the caspase 3 large subunit improved production of a biologically active IL2‐Caspase3 chimeric protein
Author(s) -
Glantz Yitav,
Sabag Ofra,
Lichtenstein Michal,
Grodzovski Inna,
LorberboumGalski Haya
Publication year - 2012
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.1515
Subject(s) - protein subunit , amino acid , fusion protein , caspase , programmed cell death , biological activity , moiety , apoptosis , biochemistry , chemistry , biology , microbiology and biotechnology , stereochemistry , recombinant dna , gene , in vitro
Designing a chimeric protein and developing a procedure for its stable production as a biologically active protein, are key steps in its potential application to clinical trails. IL2‐Caspase3 chimeric protein designed to target activated T lymphocytes was found to be a promising molecule for targeted treatment, however was found to be difficult to produce as a biological active molecule. Thus, we designed a new version of the molecule, IL2‐Caspase3s, in which six amino acids (aa 29–34) from the N ‐terminus of the large subunit of caspase 3 were excluded. Repeated expressions, productions, and partial purifications of the IL2‐Caspase3s yielded reproducible batches with consistent results. We found that IL2‐Caspase3s causes cell death in a specific, dose‐, and time‐dependent manner. Cell death due to IL2‐Caspase3s is caused by apoptosis. This improved and biologically stable IL2‐Caspase3s chimeric protein may be developed in the future for clinical trails as a promising therapy for several pathologies involving activated T‐cells. Moreover, this truncated caspase 3 sequence, lacking the N ‐terminal six amino acids of its large subunit, may be used in other caspase 3‐based chimeric proteins targeted against various human diseases, using the appropriate targeting moiety. © 2012 American Institute of Chemical Engineers Biotechnol. Prog., 2012