Optimization of binding proteins
First Claim
1. A method for manipulating nucleic acids so as to increase the binding of an encoded antibody to a particular ligand, said method comprising the steps of:
- (a) providing two or more nucleic acids encoding different antibodies all having selective binding for said particular ligand, said nucleic acids having at least one set of splicing sites, said set of splicing sites flanking opposite ends of one or more encoded CDRs, wherein the binding activity of at least one encoded antibody is known;
(b) mixing said two or more nucleic acids to produce a parent population of mixed nucleic acids encoding said antibodies;
(c) randomly incorporating said one or more encoded CDRs between said two or more nucleic acids through said set of splicing sites to produce a different population of nucleic acids than said parent population, wherein at least one antibody encoded by said different population is characterized by having increased binding to the particular ligand as compared to the antibody having the known binding activity; and
(d) identifying at least one of the antibodies having increased binding to the particular ligand as compared to the antibody having the known binding activity.
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Abstract
The invention relates to methods for manipulating nucleic acids so as to optimize the binding characteristics of an encoded binding protein by providing two or more nucleic acids encoding binding proteins having at least one set of splicing sites, the set of splicing sites flanking opposite ends of one or more encoded binding domains; mixing the nucleic acids to produce a parent population of mixed nucleic acids encoding binding proteins; and randomly incorporating the binding domains between the nucleic acids through the set of splicing sites to produce a different population of nucleic acids encoding binding proteins wherein at least one binding protein is characterized by substantially different binding characteristics than a member of the parent population.
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Citations
29 Claims
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1. A method for manipulating nucleic acids so as to increase the binding of an encoded antibody to a particular ligand, said method comprising the steps of:
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(a) providing two or more nucleic acids encoding different antibodies all having selective binding for said particular ligand, said nucleic acids having at least one set of splicing sites, said set of splicing sites flanking opposite ends of one or more encoded CDRs, wherein the binding activity of at least one encoded antibody is known; (b) mixing said two or more nucleic acids to produce a parent population of mixed nucleic acids encoding said antibodies; (c) randomly incorporating said one or more encoded CDRs between said two or more nucleic acids through said set of splicing sites to produce a different population of nucleic acids than said parent population, wherein at least one antibody encoded by said different population is characterized by having increased binding to the particular ligand as compared to the antibody having the known binding activity; and (d) identifying at least one of the antibodies having increased binding to the particular ligand as compared to the antibody having the known binding activity. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 28)
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13. A population of nucleic acids encoding antibodies, said population of nucleic acids produced by a process, comprising the steps of:
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(a) providing two or more nucleic acids encoding different antibodies all having selective binding for said particular ligand, said nucleic acids having at least one set of splicing sites, said set of splicing sites flanking opposite ends of one or more encoded CDRs, wherein the binding activity of at least one encoded antibody is known; (b) mixing said two or more nucleic acids to produce a parent population of mixed nucleic acids encoding said antibodies; and (c) randomly incorporating said one or more encoded CDRs between said two or more nucleic acids through said set of splicing sites to produce a different population of nucleic acids than said parent population, wherein at least one antibody is characterized by different binding characteristics than the antibody having the known binding activity.
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14. A population of vectors containing nucleic acids encoding antibodies, said population of vectors produced by a process, comprising the steps of:
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(a) providing two or more nucleic acids encoding different antibodies all having selective binding for said particular ligand, said nucleic acids having at least one set of splicing sites, said set of splicing sites flanking opposite ends of one or more encoded CDRs, wherein the binding activity of at least one encoded antibody is known; (b) mixing said two or more nucleic acids to produce a parent population of mixed nucleic acids encoding said antibodies; (c) randomly incorporating said one or more encoded CDRs between said two or more nucleic acids through said set of splicing sites to produce a different population of nucleic acids than said parent population, wherein at least one antibody is characterized by different binding characteristics than the antibody having the known binding activity; and (d) inserting said different population of nucleic acids encoding said antibodies into vectors. - View Dependent Claims (15)
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16. A method for manipulating nucleic acids so as to decrease the binding of an encoded antibody to a particular ligand, said method comprising the steps of:
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(a) providing two or more nucleic acids encoding different antibodies all having selective binding for said particular ligand, said nucleic acids having at least one set of splicing sites, said set of splicing sites flanking opposite ends of one or more encoded CDRs, wherein the binding activity of at least one encoded antibody is known; (b) mixing said two or more nucleic acids to produce a parent population of mixed nucleic acids encoding said antibodies; (c) randomly incorporating said one or more encoded CDRs between said two or more nucleic acids through said set of splicing sites to produce a different population of nucleic acids than said parent population, wherein at least one antibody encoded by said different population is characterized by having decreased binding to the particular ligand as compared to the antibody having the known binding activity; and (d) identifying at least one of the antibodies having decreased binding to the particular ligand as compared to the antibody having the known binding activity. - View Dependent Claims (17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 29)
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Specification