Patent ID: 8771694

Claim:
A method of designing amino acid sequences of variable domains of a humanized monoclonal antibody comprising: (a) comparing the amino acid sequences of the light and heavy chain variable domains of a monoclonal antibody to be humanized with the amino acid sequences of the light and heavy chain variable domains of two or more human antibodies; (b) selecting framework regions from a first human antibody for the light chain and from second and third human antibodies for the heavy chain based on the sequence comparison, wherein the heavy chain FR1, FR2 and FR3 are selected from the second human antibody and FR4 is selected from the third human antibody; (c) incorporating the framework regions selected in step (b) with the corresponding light and heavy chain complementarity determining regions of the monoclonal antibody to be humanized, to design humanized light and heavy chain variable domain amino acid sequences; (d) preparing DNA sequences encoding the humanized light and heavy chain variable domain amino acid sequences; (e) operably incorporating the variable domain DNA sequences into at least one vector comprising DNA sequences encoding the constant domains of the human light and heavy chain regions; (f) introducing the at least one vector into a cell; and (g) culturing the cell containing the at least one vector under conditions to produce the humanized monoclonal antibody wherein the heavy chain FR4 is selected from the human NEWM antibody, the light chain framework regions are selected from the human REI antibody, and the heavy chain FR1, FR2 and FR3 are selected from the human EU antibody.