Methods of modifying antibodies, and modified antibodies with improved functional properties

摘要

Disclosed is method of engineering an immunobinder, the immunobinder comprising a human Vk3 family light chain variable region, or fragment thereof, the Vk3 family light chain variable region comprising VL framework residues, the method comprising: A) selecting one or more amino acid positions within the Vk3 framework residues; and B) mutating the one or more amino acid positions selected for mutation, wherein the mutating comprises one or more substitutions selected from the group consisting of: wherein if the one or more light chain amino acid positions selected for mutation are of a human Vk3 family light chain variable region, the mutating comprises one or more substitutions selected from the group consisting of: (i) isoleucine (I) at amino acid position 10 using AHo or Kabat numbering system; (ii) tyrosine (Y) at amino acid position 12 using AHo or Kabat numbering system; (iii) serine (S) at amino acid position 18 using AHo or Kabat numbering system; (iv) alanine (A) at amino acid position 20 using AHo or Kabat numbering system; (v) methionine (M) at amino acid position 56 using AHo numbering system (amino acid position 48 using Kabat numbering system); (vi) valine (V) or threonine (T) at amino acid position 74 using AHo numbering system (amino acid position 58 using Kabat numbering system); (vii) asparagine (N) at amino acid position 94 using AHo numbering system (amino acid position 76 using Kabat numbering system); (viii) tyrosine (Y) or serine (S) at amino acid position 101 using AHo numbering system (amino acid position 83 using Kabat numbering system); and (ix) leucine (L) or alanine (A) at amino acid position 103 using AHo numbering (amino acid position 85 using Kabat numbering).