Bivalent, bispecific antibodies

Abstract

The present invention relates to novel domain exchanged, bivalent, bispecific antibodies, their manufacture and use.

Description

PRIORITY TO RELATED APPLICATION(S)[0001]This application claims the benefit of European Patent Application No. 07024864.6, filed Dec. 21, 2007, which is hereby incorporated by reference in its entirety.[0002]The present invention relates to novel bivalent, bispecific antibodies, their manufacture and use.BACKGROUND OF THE INVENTION[0003]Engineered proteins, such as bi- or multispecific antibodies capable of binding two or more antigens are known in the art. Such multispecific binding proteins can be generated using cell fusion, chemical conjugation, or recombinant DNA techniques.[0004]A wide variety of recombinant bispecific antibody formats have been developed in the recent past, e.g. tetravalent bispecific antibodies by fusion of, e.g. an IgG antibody format and single chain domains (see e.g. Morrison, S. L., et al, Nature Biotech 15 (1997) 159-163; WO2001077342; and Coloma, M. J., Nature Biotech 25 (2007) 1233-1234).[0005]Also several other new formats wherein the antibody core s...

AI Technical Summary

Benefits of technology

[0043]The CH3 domains of said bivalent, bispecific antibody according to the invention can be altered by the “knob-into-holes” technology which described with in detail with several examples in e.g. WO96 / 027011, Ridgway J B, et al, Protein Eng 9 (1996) 617-621; and Merchant A. M., et al, Nat Biotechnol 16 (1998) 677-681. In this method the interaction surfaces of the two CH3 domains are altered to increase the heterodimerisation of both heavy chains containing these two CH3 domains. Each of the two CH3 domains (of the two heavy chains) can be the “knob”, while the other is the “hole”. The introduction of a disulfide bridge stabilizes the heterodimers (Merchant A. M, et al, Nature Biotech 16 (1998) 677-681; Atwell S, Ridgway J B, Wells J A, Carter P., J Mol Biol 270 (1997) 26-35) and increases the yield.

[0090]Amino acid sequence variants (or mutants) of the bivalent, bispecific antibody are prepared by introducing appropriate nucleotide changes into the antibody DNA, or by nucleotide synthesis. Such modifications can be performed, however, only in a very limited range, e.g. as described above. For example, the modifications do not alter the above mentioned antibody characteristics such as the IgG isotype and antigen binding, but may improve the yield of the recombinant production, protein stability or facilitate the purification.

Problems solved by technology

While it is obvious that linkers have advantages for the engineering of bispecific antibodies, they may also cause problems in therapeutic settings.

Further more, the flexible nature of these peptides makes them more prone to proteolytic cleavage, potentially leading to poor antibody stability, aggregation and increased immunogenicity.

In general the same problem of mispaired byproducts remains if recombinant expression techniques are used.

Although this format appears very attractive, no data describing progression towards the clinic are currently available.

Thus this technique is not appropriate for easily developing recombinant, bivalent, bispecific antibodies against two antigens starting from two antibodies against the first and the second antigen, as either the heavy chains of these antibodies an / or the identical light chains have to be optimized.

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