Patent ID: 9175087
Filing Date: 2015-11-03
Classification: A61K,A61P,C07K,G16B

Abstract:
1. A method of making a humanized antibody or fragment thereof, comprising: (a) identifying a human variable region homologous to a variable region of a non-human antibody; (b) building a homology model of the variable region of the non-human antibody using human variable region identified in step (a) and running molecular dynamic simulations to obtain molecular dynamic trajectory of the variable region of the non-human antibody; (c) analyzing the molecular dynamic trajectory of the variable region of the non-human antibody and determining amino acid flexibility scores to identify amino acids in the variable region of the non-human antibody which are flexible and amino acids that flank the flexible residue; (d) building homology models of human antibody germ lines and running molecular dynamic simulations to obtain molecular dynamic trajectories of the variable region of the human antibody germ lines; (e) analyzing the molecular dynamic trajectories of the variable region of the human antibody germ lines and determining amino acid flexibility scores to identify amino acids in the variable region of the human antibody germ lines which are flexible and amino acids that flank the flexible residue; (f) identifying the human variable region of the human antibody germ lines that displays molecular dynamic trajectory most similar to the molecular dynamic trajectory of the variable region of the non-human antibody and identifying humanizing mutations by comparing the flexibility scores for the most flexible amino acids of steps (c) and (e); (g) replacing the identified amino acids of step (c) with the identified amino acids of step (f) to produce a humanized variable region; (h) confirming that the humanized variable region of step (g) resembles a human antibody by comparing the humanized variable region molecular dynamic trajectory with the molecular dynamic trajectory of the variable region of the human antibody germ lines; and (i) recombinantly constructing a humanized variable region by joining the humanized variable region of step (g) with a human sequence to yield a humanized antibody or fragment thereof.