Patent ID: 8685897

Claim:
A method for preparing a humanized antibody having a desired property, which method comprises: (a) contacting one or more host cells that express or can be induced to express Activation Induced Cytidine Deaminase (AID) with: (1) a first nucleic acid sequence which encodes one or more human antibody heavy chain protein scaffolds, wherein each heavy chain protein scaffold encodes all of a complementarity determining region 1 (CDR1) and all of a complementarity determining region 2 (CDR2) from the human antibody, (2) a first library of nucleic acid sequences, wherein each nucleic acid sequence of the library encodes all of a complementarity determining region 3 (CDR3) from a non-human antibody heavy chain protein, and wherein the first nucleic acid sequence of (i) and the first library of (ii) are operably linked in a first expression vector to produce a library of humanized antibody heavy chain protein-encoding polynucleotides comprising the CDR1 and CDR2 of the first nucleic acid sequence and the CDR3 of the first library, (3) a second nucleic acid sequence which encodes one or more human antibody light chain protein scaffolds, wherein each light chain protein scaffold encodes all of a CDR1 and all of a CDR2 from the human antibody, and (4) a second library of nucleic acid sequences, wherein each nucleic acid sequence of the library encodes all of a CDR3 from a non-human antibody light chain protein, and wherein the second nucleic acid sequence of (iii) and the second library of (iv) are operably linked in a second expression vector to produce a library of humanized antibody light chain protein-encoding polynucleotides comprising the CDR1 and CDR2 of the second nucleic acid sequence and the CDR3 of the second library, (b) whereupon AID expression in the one or more host cells induces one or more mutations in the nucleic acid sequences of the library of humanized antibody heavy chain protein-encoding polynucleotides produced in (a)(2) and one or more mutations in the nucleic acid sequences of the library of humanized antibody light chain protein-encoding polynucleotides produced in (a)(4), whereupon a humanized antibody having a desired property is produced by the one or more host cells, and (c) identifying or isolating the humanized antibody having the desired property.