Method for generating high affinity antibodies
OwnerSanjaya Singh, Catherine Foster +1inventors
InventorsSanjaya Singh, Catherine Foster, Herren Wu
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Abstract
The invention relates to high affinity human monoclonal antibodies, particularly those directed against isotypic determinants of immunoglobulin E (IgE), as well as direct equivalents and derivatives of these antibodies. These antibodies bind to their respective target with an affinity at least 100 fold greater than the original parent antibody. These antibodies are useful for diagnostics, prophylaxis and treatment of disease.
Claims29
A method for generating a humanized, high affinity antibody from an antibody of interest comprising the steps of:
- a. obtaining an antibody of interest (“parent”) comprising a heavy and light chain variable domain;
- b. determining the amino acid sequence of the heavy (H) and light (L) chain variable domains of the antibody of step (a);
- c. selecting a suitable human template as the framework for the H and L chain variable domains of the high affinity antibody to be made;
- d. generating a library of random amino acid substitutions in the parent CDRs and/or framework (FR) regions chosen in step (c);
- e. constructing an expression vector comprising the H and L chain variable domain variants generated in step (d);
- f. screening the library for variants with increased binding affinity as compared to the parent antibody.
A method for generating a high affinity antibody from an antibody of interest comprising the steps of:
- a. obtaining an antibody of interest (“parent”) comprising a heavy and light chain variable domains;
- b. determining the amino acid sequence of the heavy (H) and light (L) chain variable domains of the antibody of step (a);
- c. selecting a suitable template as the framework for the H and L chain variable domains of the high affinity antibody to be made;
- d. generating a library of random amino acid substitutions in the parent CDRs and/or framework (FR) regions chosen in step (c);
- e. constructing an expression vector comprising the H chain variants generated in step (d) and an expression vector comrising the light chain variants generated in step (d);
- f. expressing the heavy and light chain variants of step (e);
- g. screening the library for variants having increased binding affinity as compared to the parent antibody.
A high affinity antibody, wherein the binding affinity is at least 100 fold higher than the parent antibody from which it was generated.
A high affinity antibody, wherein the binding affinity is at least 1000 fold higher than the parent antibody from which it was generated.
Dependencies are inferred from the claim text.