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US 2003/0166871 A1Published application
Humanization of murine antibody
AssigneeThe Scripps Research Institute
InventorsCarlos F. Barbas III, Christoph Rader
Life of this patent
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Abstract
A humanized murine antibody is provided. The amino acid sequences of a light chain complementarity determining region from a mouse antibody are grafted onto a human light chain, and a heavy chain complementarity determining region from a mouse antibody are grafted onto a human antibody heavy chain to produce libraries from which a humanized murine antibody having the desired specificity is selected.
Claims15
A method of producing a humanized mouse monoclonal antibody which consists of the steps:
- (a) constructing a first human antibody heavy or light chain library in which each chain has at least one complementarity determining region (CDR) loop and each such loop is flanked by unaltered human framework residues and has the amino acid sequence of a corresponding mouse antibody heavy or light chain CDR loop;
- (b) constructing a second human antibody heavy or light chain library, in which each complementarity determining region (CDR) loop and each such loop is flanked by unaltered human framework residues and has the amino acid sequence of a corresponding mouse antibody heavy or light chain CDR loop, wherein the chains of said second library are the complementary heavy or light chain of said first library chains, such that one library is a light chain library and the other library is a heavy chain library;
- (c) creating a library of heavy and light chain pairs by combining chains from said library of step (a) with a complementary chain from an antibody which binds a preselected antigen forming a library of heavy and light chain pairs;
- (d) isolating antigen binding chains of step (a) by selecting from the library of step (c), a heavy and light chain pair which binds to said preselected antigen, isolating from the antigen binding heavy and light chain pair, the antigen binding chains of step (a);
- (e) creating a humanized pair library by combining isolated antigen binding chains of step (d), with chains of said library of step (b), so that a first isolated chain of step (d), together with a complementary second chain in said library of step (b) combine to form a heavy and light chain humanized pair library;
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- (f) selecting from the humanized pair library of step (e) a humanized heavy and light chain pair that binds to said preselected antigen;
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- (g) combining two of said selected humanized heavy and light chain pair of step (f) to form a whole antibody.
6 more elements
A method of producing a humanized mouse monoclonal antibody heavy and light chain pair which consists of the steps:
- (a) constructing a human light chain library wherein each said light chain has at least one complementarity determining region (CDR) loop that has an amino acid sequence corresponding to that of a mouse light chain CDR loop and each such loop is flanked by unaltered human framework residues;
- (b) selecting a light chain from the light chain library of step (a), wherein the selection comprises combining a heavy chain from an antibody that binds a preselected antigen with a light chain from the library of step (a) to form a heavy and light chain pair library, screening said library for binding to said preselected antigen, and isolating the light chains that bind to said preselected antigen;
- (c) constructing a library of human heavy chains wherein each said heavy chain has at least one CDR loop that has an amino acid sequence corresponding to a mouse heavy chain CDR loop and each such loop is flanked by unaltered human framework residues;
- (d) combining a chain from the heavy chain library of step (c) with a selected chain of step (b) to produce a second library of human heavy and light chain pairs;
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- (e) selecting an antigen binding human heavy and light chain pair from the human library of step (d) by screening said library for binding with said preselected antigen and selecting those heavy and light chain pairs that bind said preselected antigen.
3 more elements
Dependencies are inferred from the claim text.