Fab molecules with a rodent hinge region and a non-rodent CH1 region
The disclosure relates to novel Fab molecules comprising a modified heavy chain constant region. The modified constant region prevents the recognition of the Fab molecules by anti-Fab antibodies present in a host's serum. The disclosure further relates to methods of generating such modified Fab molecules for biological, diagnostic, pharmaceutical and other uses.
1. A Fab comprising a modified heavy chain constant region, wherein said modified heavy chain constant region comprises a CH1 domain and a hinge region, wherein the hinge region is of a rodent IgG isotype and consists of the amino acid sequence of any one of ERRNGGIGHKC (SEQ ID NO: 32), EERNGGIGHKC (SEQ ID NO: 93), ERRQGGIGHKC (SEQ ID NO: 34) or VPREC (SEQ ID NO: 26) and wherein the CH1 domain is a wildtype human IgG1 CH1 domain or any allotype of a wildtype human IgG1 CH1 domain.
2. The Fab of claim 1 , wherein the CH1 domain comprises at least the amino acid sequence of any one of DKKV (SEQ ID NO: 86), DKRV (SEQ ID NO: 87), EKKV (SEQ ID NO: 88), DKKI (SEQ ID NO: 89), DKGV (SEQ ID NO: 90) or DKEV (SEQ ID NO: 91) from position 212 to 215 according to EU numbering.
3. The Fab of claim 1 , wherein the Fab CH1 domain consists of the amino acid sequence of any one of
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKV (SEQ ID NO: 1),
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRV (SEQ ID NO: 2),
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKI (SEQ ID NO: 7),
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVEKKV (SEQ ID NO: 8),
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKGV (SEQ ID NO: 9) or
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKEV (SEQ ID NO: 10.
4. The Fab of claim 1 , wherein the CH1 domain comprises the amino acid substitution. D212E, K214G, K214E, R214G, R214G or V215I.
5. The Fab of claim 1 , comprising a modified heavy chain constant region comprising a CH1 domain and a hinge region in order from N- to C-terminus, and wherein the CH1 domain consists of an amino acid sequence of any one of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 7-10.
6. The Fab of claim 1 , comprising a modified heavy chain constant region consisting of an amino acid sequence selected from the group consisting of SEQ ID NO: 70-76.
7. The Fab of claim 1 , comprising a modified heavy chain constant region comprising a CH1 domain and a hinge region in order from N- to C-terminus, and wherein the CH1 domain consists of the amino acid sequence
ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKEV (SEQ ID NO: 10) and the hinge region consists of the amino acid sequence ERRQGGIGHKC (SEQ ID NO: 34).