Absolute Antibody offers a royalty free antibody humanization service for antibodies from any species. Our antibody humanization service consists of the following:
- Sequencing of your hybridoma if required (for more information see our hybridoma sequencing service)
- Production of wild type and chimeric controls (0.2 mg)
- Detailed analysis of your antibody sequence, CDR identification and comparison of framework regions to human germline antibody sequences
- In silico humanization followed by expression and purification of 3-5 humanized variants for confirmation of activity compared to controls
- A report containing details of humanization strategy and the final humanized sequence(s)
Antibody humanization background information
The strive to reduce the immunogenicity of antibodies in the 1980s first led to the engineering of chimeric antibodies (1,2), consisting of human constant domains with the original non-human variable domains. Shortly after, this was followed by humanization (3) where complementarity determining regions (CDRs) from the original antibody were grafted on to a human variable domain framework. Crystal structures subsequently showed that selection of the appropriate human framework supports the CDRs in the correct orientation and minimizes affinity losses (4). Although the process of humanization, also referred to as antibody reshaping or germlining, has developed since this early work the concept remains much the same – namely to make the antibody more human-like and less immunogenic. It is now routine for all therapeutic antibodies originally derived from non-human sources (e.g. hybridomas) to undergo humanization as part of the antibody development pipeline. For more information on the background to humanization please refer to the Absolute Antibody resource section on humanization.
- Boulianne, G.L., Hozumi, N., and Shulman, M.J. (1984). Production of functional chimaeric mouse/human antibody. Nature 312, 643–646.
- Morrison, S.L., Johnson, M.J., Herzenberg, L.A., and Oi, V.T. (1984). Chimeric human antibody molecules: mouse antigen-binding domains with human constant region domains. Proc. Natl. Acad. Sci. U.S.A. 81, 6851–6855.
- Jones, P.T., Dear, P.H., Foote, J., Neuberger, M.S., and Winter, G. (1986). Replacing the complementarity-determining regions in a human antibody with those from a mouse. Nature 321, 522–525.
- Cheetham, G.M., Hale, G., Waldmann, H., and Bloomer, A.C. (1998). Crystal structures of a rat anti-CD52 (CAMPATH-1) therapeutic antibody Fab fragment and its humanized counterpart. J. Mol. Biol. 284, 85–99.