Production of biologically active IgG hinge-tag soluble epidermal growth factor receptors (ErbB).
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.
Otani
Takayuki
T
Department of Medical and Bioengineering Science, Graduate School of Natural Science and Technology, Okayama University, 3.1.1 Tsushima-Naka, Kita-ku, Okayama, 700-8530, Japan.
Hashizume
Toshihiro
T
Nagaoka
Tadahiro
T
Fukuda
Tomoko
T
Tang
Careen K
CK
Salomon
David S
DS
Seno
Masaharu
M
D000818:Animals; D066259:Betacellulin; D001709:Biotechnology / Q000379:methods*; D019556:COS Cells; D002522:Chlorocebus aethiops; D066246:ErbB Receptors / Q000096:biosynthesis*; D006801:Humans; D007118:Immunoassay; D007074:Immunoglobulin G / Q000096:biosynthesis*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D007700:Kinetics; D008970:Molecular Weight; D020890:Neuregulin-1 / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis*; D012995:Solubility
C587851:BTC protein, human; D066259:Betacellulin; D007074:Immunoglobulin G; D036341:Intercellular Signaling Peptides and Proteins; D020890:Neuregulin-1; D011994:Recombinant Proteins; D066246:ErbB Receptors