Abstract:
PROBLEM TO BE SOLVED: To provide novel methods for making and refolding insoluble or aggregated proteins having free cysteine. SOLUTION: The present invention relates to novel methods for making and refolding insoluble or aggregated proteins having free cysteines in which a host cell expressing the protein is exposed to a cysteine blocking agent. The soluble, refolded proteins produced by the novel methods can then be modified to increase their effectiveness. Such modifications include attaching a PEG moiety to form PEGylated proteins. COPYRIGHT: (C)2011,JPO&INPIT
Abstract:
PROBLEM TO BE SOLVED: To provide a method of obtaining soluble proteins having at least one added free cysteines.SOLUTION: The method comprises the steps of: (a) obtaining a host cell capable of expressing the soluble protein having at least one added free cysteines; (b) exposing the host cell to a cysteine blocking agent forming a stable mixed disulfide with the at least one added free cysteines before the step (c); and (c) isolating the soluble protein from the host cell. The produced soluble proteins can be modified to increase their effectiveness, and such modifications include attaching a PEG moiety to form pegylated proteins.
Abstract:
PROBLEM TO BE SOLVED: To provide an isolated monoPEGylated growth hormone protein. SOLUTION: The isolated monoPEGylated growth hormone protein comprises a polyethylene glycol attached to an amino acid selected from the group consisting of amino acid 2, 3, 5, 38, 40, 55, 57, 99, 101, 102, 103, 132, 133, 134, 137, 140, 141, 143, 144, 147, 148, 149, 186, and 187 of growth hormone, wherein the obtained monoPEGylated growth hormone protein stimulates proliferation in vitro of a cell line that proliferates in response to growth hormone. COPYRIGHT: (C)2011,JPO&INPIT
Abstract:
PROBLEM TO BE SOLVED: To provide a preferred site for adding a cysteine residue to the growth hormone supergene family comprising greater than 20 structurally related cytokine and growth factors, or for introducing cysteine substitution, and a protein and protein derivatives produced thereby. SOLUTION: A general method for preparing a site-specific biologically active conjugate of the growth hormone supergene family proteins such as β-interferon or the like is provided. This method comprises a step in which a cysteine residue is added to a non-essential region of this protein, or a step in which a non-essential amino acid in the protein is substituted with a cysteine residue by using site-specific mutagenesis, and then a step in which the protein is connected to a cysteine reactive polymer through the added cysteine residue or another type cysteine reactive part is connected with a covalent bond. COPYRIGHT: (C)2009,JPO&INPIT