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US 9,803,193 B2Granted
Protein screening methods
AssigneeX-Body, Inc.
InventorsRichard W. Wagner, Alexander Litovchick, Yan Chen
Life of this patent
Estimating the term…
Abstract
The invention provides methods and compositions useful for identifying polypeptides with desired characteristics in vitro.
Claims44
A method of selecting an isolated nucleic acid molecule encoding a polypeptide capable of binding to an antigen of interest, comprising the steps of:
- (a) providing a library of nucleic acid-polypeptide complexes, wherein at least a portion of the complexes contain different polypeptide-encoding sequences;
- (b) contacting the library with an antigen of interest;
- (c) selecting from the library at least one nucleic acid-polypeptide complex that binds to the antigen of interest;
- and
- (d) isolating the polypeptide-encoding sequence of the selected nucleic acid-polypeptide complex,
- wherein each member of the library of nucleic acid-polypeptide complexes comprises:
- (a) a first nucleic acid molecule comprising a polypeptide-encoding sequence;
- (b) a polypeptide encoded by the first nucleic acid molecule;
- and
- (c) a second nucleic acid molecule comprising a nucleic sequence complementary to a portion of the first nucleic acid molecule, wherein the first nucleic acid molecule is bound to the second nucleic acid molecule through complementary nucleic acid base pairing, and wherein the second nucleic acid molecule is non-covalently bound to the polypeptide.
7 more elements
A method of producing a polypeptide capable of binding to an antigen of interest, comprising:
- (a) providing a polypeptide-encoding sequence selected by;
- (i) providing a library of nucleic acid-polypeptide complexes, wherein at least a portion of the complexes contain different polypeptide-encoding sequences;
- (ii) contacting the library with an antigen of interest;
- (iii) selecting from the library at least one nucleic acid-polypeptide complex that binds to the antigen of interest;
- and
- (iv) isolating the polypeptide-encoding sequence of the selected nucleic acid-polypeptide complex,
- wherein each member of the library of nucleic acid-polypeptide complexes comprises:
- (i) a first nucleic acid molecule comprising a polypeptide-encoding sequence;
- (ii) a polypeptide encoded by the first nucleic acid molecule;
- and
- (iii) a second nucleic acid molecule comprising a nucleic sequence complementary to a portion of the first nucleic acid molecule, wherein the first nucleic acid molecule is bound to the second nucleic acid molecule through complementary nucleic acid base pairing, and wherein the second nucleic acid molecule is non-covalently bound to the polypeptide;
- and
- (b) expressing the polypeptide encoded by the polypeptide-encoding sequence.
10 more elements
A method of producing a polypeptide capable of binding to an antigen of interest, comprising:
- (a) providing a polypeptide-encoding sequence selected by
- (i) providing a library of nucleic acid-polypeptide complexes, wherein at least a portion of the complexes contain different polypeptide-encoding sequences;
- (ii) contacting the library with an antigen of interest;
- (iii) selecting from the library at least one nucleic acid-polypeptide complex that binds to the antigen of interest;
- and
- (iv) isolating the polypeptide-encoding sequence of the selected nucleic acid-polypeptide complex,
- wherein each member of the library of nucleic acid-polypeptide complexes comprises:
- (i) a nucleic acid molecule comprising a polypeptide-encoding sequence, wherein the nucleic acid molecule is covalently bound to a first high affinity ligand;
- (ii) a polypeptide encoded by the nucleic acid molecule, wherein the polypeptide is covalently linked at the C-terminal residue through a peptide acceptor to a second high affinity ligand;
- and
- (iii) a single multimeric ligand acceptor protein, wherein the single multimeric ligand acceptor protein is bound noncovalently and directly to both the first and second high affinity ligand at distinct ligand binding sites on the ligand acceptor protein, thereby noncovalently linking the nucleic acid molecule and the polypeptide;
- and
- (b) expressing the polypeptide encoded by the polypeptide-encoding sequence.
10 more elements
A method of producing a polypeptide capable of binding to an antigen of interest, comprising:
- (a) providing a polypeptide-encoding sequence selected by
- (i) providing a library of nucleic acid-polypeptide complexes, wherein at least a portion of the complexes contain different polypeptide-encoding sequences;
- (ii) contacting the library with an antigen of interest;
- (iii) selecting from the library at least one nucleic acid-polypeptide complex that binds to the antigen of interest;
- and
- (iv) isolating the polypeptide-encoding sequence of the selected nucleic acid-polypeptide complex,
- wherein each member of the library of nucleic acid-polypeptide complexes comprises:
- (i) a nucleic acid molecule comprising a polypeptide-encoding sequence, wherein the 3′ region of the nucleic acid molecule is covalently bound to a first biotin molecule;
- (ii) a polypeptide encoded by the nucleic acid molecule, wherein the polypeptide is covalently linked at the C-terminal residue through a peptide acceptor to a second biotin molecule;
- and
- (iii) a single multimeric biotin-binding protein, wherein the biotin-binding protein is bound noncovalently and directly to both the first and second biotin molecules at distinct ligand binding sites on the biotin-binding protein, thereby noncovalently linking the nucleic acid molecule and the polypeptide;
- and
- (b) expressing the polypeptide encoded by the polypeptide-encoding sequence.
10 more elements
A method of selecting an isolated nucleic acid molecule encoding a polypeptide capable of binding to an antigen of interest, comprising the steps of:
- (i) providing a library of nucleic acid-polypeptide complexes, wherein at least a portion of the complexes contain different polypeptide-encoding sequences;
- (ii) contacting the library with an antigen of interest;
- (iii) selecting from the library at least one nucleic acid-polypeptide complex that binds to the antigen of interest;
- and
- (iv) isolating the polypeptide-encoding sequence of the selected nucleic acid-polypeptide complex,
- wherein each member of the library of nucleic acid-polypeptide complexes comprises:
- (i) a nucleic acid molecule comprising a polypeptide-encoding sequence, wherein the nucleic acid molecule is covalently bound to a first high affinity ligand;
- (ii) a polypeptide encoded by the nucleic acid molecule, wherein the polypeptide is covalently linked at the C-terminal residue through a peptide acceptor to a second high affinity ligand;
- and
- (iii) a single multimeric ligand acceptor protein, wherein the single multimeric ligand acceptor protein is bound noncovalently and directly to both the first and second high affinity ligand at distinct ligand binding sites on the ligand acceptor protein, thereby noncovalently linking the nucleic acid molecule and the polypeptide.
7 more elements
A method of selecting an isolated nucleic acid molecule encoding a polypeptide capable of binding to an antigen of interest, comprising the steps of:
- (a) providing a library of nucleic acid-polypeptide complexes, wherein at least a portion of the complexes contain different polypeptide-encoding sequences;
- (b) contacting the library with an antigen of interest;
- (c) selecting from the library at least one nucleic acid-polypeptide complex that binds to the antigen of interest;
- and
- (d) isolating the polypeptide-encoding sequence of the selected nucleic acid-polypeptide complex,
- wherein each member of the library of nucleic acid-polypeptide complexes comprises:
- (a) a nucleic acid molecule comprising a polypeptide-encoding sequence, wherein the 3′ region of the nucleic acid molecule is covalently bound to a first biotin molecule;
- (b) a polypeptide encoded by the nucleic acid molecule, wherein the polypeptide is covalently linked at the C-terminal residue through a peptide acceptor to a second biotin molecule;
- and
- (c) a single multimeric biotin-binding protein, wherein the biotin-binding protein is bound noncovalently and directly to both the first and second biotin molecules at distinct ligand binding sites on the biotin-binding protein, thereby noncovalently linking the nucleic acid molecule and the polypeptide.
7 more elements
Dependencies are inferred from the claim text.