Cdr-repaired antibodies
AssigneeGenentech, Inc.
InventorMark S. Dennis
Life of this patent
Estimating the term…
Abstract
The present application concerns restoring antigen binding during humanization of antibodies through the selection of repaired hypervariable regions rather than through framework changes.
Claims38
An altered antibody which binds an antigen with a binding affinity (Kd) value of no more than about 5×10−7 M, the altered antibody comprising variable heavy (VH) and variable light (VL) acceptor human frameworks and one or more altered hypervariable regions derived from a non-human antibody which binds the antigen, wherein the VH and VL frameworks lack human to non-human amino acid substitutions therein.
A method of making an altered antibody comprising incorporating non-human hypervariable region residues into an acceptor human framework and further comprising introducing one or more amino acid substitutions in one or more hypervariable regions, without modifying the acceptor human framework sequence, and selecting an antibody with a binding affinity (Kd) value of no more than about 5×10−7 M.
A method of mutating a nucleic acid sequence comprising:
- (a) annealing from about two to about 20 oligonucleotides to a single stranded nucleic acid template, wherein the oligonucleotide:template ratio for each oligonucleotide is greater than 1;
- (b) removing excess unannealed oligonucleotides;
- and
- (c) filling in a nucleic acid strand which is complementary to the template.
A method of selecting an altered antibody comprising:
- (a) preparing nucleic acid encoding at least the variable heavy (VH) and variable light (VL) domains of an antibody, each comprising an acceptor human framework and hypervariable regions of a non-human antibody;
- (b) substituting hypervariable region residues by introducing an approximately 10-50 percent mutation rate into the nucleic acid so as to maintain a bias towards the non-human hypervariable region sequences;
- and
- (c) selecting one or more altered antibodies that bind antigen.
Dependencies are inferred from the claim text.