TY - JOUR
T1 - Production of biologically active IgG hinge-tag soluble epidermal growth factor receptors (ErbB)
AU - Otani, Takayuki
AU - Hashizume, Toshihiro
AU - Nagaoka, Tadahiro
AU - Fukuda, Tomoko
AU - Tang, Careen K.
AU - Salomon, David S.
AU - Seno, Masaharu
N1 - Funding Information:
Acknowledgements We thank Ms. N. Hironaka for the construction of sErbB2 and sErbB3 expression plasmid. This research was partly supported by the Ministry of Education, Culture, Sports, Science and Technology Grant-in-Aid for Scientific Research (B) Nos. 18300164 and 21300179.
PY - 2010/2
Y1 - 2010/2
N2 - The extracellular domains (ECD) of epidermal growth factor receptors, ErbB1, 2, 3 and 4, were designed as soluble dimeric forms. Each ECD was fused to a short hinge region derived from IgG, such that the stable dimer could be formed with disulfide bridges. This hinge-tagged design minimized the molecular weight to approximately 50% of the conventional Fc-fusion design without an Fc domain of IgG. The refolded dimers could be easily analyzed and characterized by SDS-PAGE. Hinge-tagged soluble ErbBs demonstrated significant affinity for betacellulin and heregulin. The IgG hinge-tag should be a simple method to design soluble dimers that would be useful for high throughput screening of ligands, antagonists or derivatives.
AB - The extracellular domains (ECD) of epidermal growth factor receptors, ErbB1, 2, 3 and 4, were designed as soluble dimeric forms. Each ECD was fused to a short hinge region derived from IgG, such that the stable dimer could be formed with disulfide bridges. This hinge-tagged design minimized the molecular weight to approximately 50% of the conventional Fc-fusion design without an Fc domain of IgG. The refolded dimers could be easily analyzed and characterized by SDS-PAGE. Hinge-tagged soluble ErbBs demonstrated significant affinity for betacellulin and heregulin. The IgG hinge-tag should be a simple method to design soluble dimers that would be useful for high throughput screening of ligands, antagonists or derivatives.
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U2 - 10.1007/s10529-009-0160-9
DO - 10.1007/s10529-009-0160-9
M3 - Article
C2 - 19898750
AN - SCOPUS:77649338416
VL - 32
SP - 361
EP - 366
JO - Biotechnology Letters
JF - Biotechnology Letters
SN - 0141-5492
IS - 3
ER -