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

摘要

Disclosed is a method of engineering an immunobinder, the immunobinder comprising a human VH1a family heavy chain variable region, or fragment thereof, the VH1a heavy chain variable region comprising VH framework residues, the method comprising: A) selecting one or more amino acid positions within the VH1a 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: (i) glutamic acid (E) at amino acid position 1 using AHo or Kabat numbering system; (ii) glutamic acid (E) at amino acid position 6 using AHo or Kabat numbering system; (iii) methionine (M) at amino acid position 13 using AHo numbering system (amino acid position 12 using Kabat numbering system): (iv) glutamic acid (E) or glutamine (Q) at amino acid position 14 using AHo numbering system (amino acid position 13 using Kabat numbering system); (v) leucine (L) at amino acid position 19 using AHo numbering system (amino acid position 18 using Kabat numbering system); (vi) isoleucine (I) at amino acid position 21 using AHo numbering system (amino acid position 20 using Kabat numbering system); (vii) phenylalanine (F), serine (S), histidine (H) or aspartic acid (D) at amino acid position 90 using AHo numbering system (amino acid position 79 using Kabat numbering system); (viii) aspartic acid (D) or glutamine (Q) at amino acid position 92 using AHo numbering system (amino acid position 81 using Kabat numbering system); (ix) glycine (G), asparagine (N) or threonine (T) at amino acid position 95 using AHo numbering system (amino acid position 82b using Kabat numbering system); and (x) threonine (T), alanine (A), proline (P) or phenylalanine (F) at amino acid position 98 using AHo numbering (amino acid position 84 using Kabat numbering.