Trimeric collagen scaffold antibodies
AssigneeIndustrial Technology Research Institute
InventorsMin-Yuan Chou, Chia-Yu Fan, Chuan-Chuan Huang, Hsiu-Chuan Li
Life of this patent
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Abstract
A collagen scaffold domain, including a collagenous or collagen-like domain, which directs self-trimerization is provided. The collagen scaffold domain can be fused to one or more heterologous domains, such as an antibody domain. Methods for generating and using the scaffold domains and fusion proteins are also provided.
Claims33
A trimeric soluble antibody comprising three polypeptides, wherein each polypeptide comprises:
- (a) a collagen scaffold domain comprising at least 10 G-X-Y repeats;
- wherein G is glycine, X is any amino acid, and Y is any amino acid,
- wherein at least 10 of the G-X-Y repeats are G-P-P or G-P-O,
- wherein at least 6 of the G-X-Y repeats are G-P-O, and
- wherein P is proline and O is hydroxyproline;
- and
- (b) an antibody domain;
- wherein the collagen scaffold domains of the three polypeptides interact with each other to form a trimeric soluble antibody that specifically binds to a ligand with an avidity of at least 107 M−1.
A method for generating a trimeric soluble antibody comprising:
- (a) joining a nucleic acid encoding a collagen scaffold domain comprising 10-30 G-X-Y repeats;
- wherein G is glycine, X is any amino acid, and Y is any amino acid, and
- wherein at least 10 of the G-X-Y repeats are G-P-P;
- in-frame with a nucleic acid encoding an antibody domain for a ligand;
- and
- (b) expressing the encoded polypeptide in a cell that hydroxyprolinates at least 6 of the G-P-P repeats at the Y position;
- wherein the hydroxyprolinated collagen scaffold domains of three polypeptides interact with each other to form a trimeric soluble antibody that specifically binds to a ligand with an avidity of at least 107 M−1.
A method for modulating the biological activity of a ligand comprising incubating a trimeric soluble antibody comprising three polypeptides with the ligand;
- wherein each polypeptide comprises:
- (a) a collagen scaffold domain comprising at least 10 G-X-Y repeats;
- wherein G is glycine, X is any amino acid, and Y is any amino acid,
- wherein at least 10 of the G-X-Y repeats are G-P-P or G-P-O,
- wherein at least 6 of the G-X-Y repeats are G-P-O, and
- wherein P is proline and O is hydroxyproline;
- and
- (b) an antibody domain;
- wherein the collagen scaffold domains of three polypeptides interact with each other to form a trimeric soluble antibody that specifically binds to a ligand with an avidity of at least 107 M−1;
- and
- wherein the binding of the trimeric soluble antibody to the ligand modulates the biological activity of the ligand.
A method for detecting a ligand comprising
- (1) incubating a trimeric soluble antibody comprising three polypeptides with the ligand;
- wherein each polypeptide comprises:
- (a) a collagen scaffold domain comprising at least 10 G-X-Y repeats;
- wherein G is glycine, X is any amino acid, and Y is any amino acid,
- wherein at least 10 of the G-X-Y repeats are G-P-P or G-P-O,
- wherein at least 6 of the G-X-Y repeats are G-P-O, and
- wherein P is proline and O is hydroxyproline;
- and
- (b) an antibody domain;
- wherein the collagen scaffold domains of three polypeptides interact with each other to form a trimeric soluble antibody that specifically binds to a ligand with an avidity of at least 107 M−1;
- and
- (2) detecting the binding of the trimeric soluble antibody to the ligand.
A trimeric soluble antibody comprising three polypeptides, wherein each polypeptide comprises:
- (a) a collagen scaffold domain comprising at least 10 G-X-Y repeats;
- wherein G is glycine, X is any amino acid, and Y is any amino acid;
- wherein at least 6 of the Y residues are hydroxyproline;
- and
- (b) an antibody domain;
- wherein the collagen scaffold domains of the three polypeptides interact with each other to form a trimeric soluble antibody that specifically binds to a ligand with an avidity of at least 107 M−1.
A trimeric soluble antibody comprising three polypeptides, wherein each polypeptide comprises:
- (a) a collagen scaffold domain comprising at least 6 G-P-O repeats;
- wherein G is glycine, P is proline, and O is hydroxyproline;
- and
- (b) an antibody domain;
- wherein the collagen scaffold domains of the three polypeptides interact with each other to form a trimeric soluble antibody that specifically binds to a ligand with an avidity of at least 107 M−1.
Dependencies are inferred from the claim text.