| 主权项 |
1. A method of identifying a variant of an antibody with enhanced antigen binding affinity, the method comprising:
(a) determining a three-dimensional representation of an antibody-antigen interface of a first antibody, (b) conformationally sampling point mutations of amino acid residues of the antibody at the antibody-antigen interface, (c) selecting the point mutations of step (b) that are conformationally allowed, (d) selecting the point mutations of step (c) that have both a Boltzmann averaged predictor based on a change of a change of a free energy of binding (ΔΔG*) of less than zero kcal/mol and a Boltzmann averaged predictor based on only a change of a change of the polar component of a free energy of binding (ΔΔEpol*) of less than zero kcal/mol, and (e) creating a focused library of antibodies containing at least one point mutation from the point mutations of step (d), (f) screening the antibodies of step (e) for enhanced antigen-binding affinity in vitro; and (g) selecting an antibody of step (f) having enhanced binding affinity relative to the first antibody, thereby identifying a variant of an antibody with enhanced antigen binding affinity. |
