Monoclonal antibodies to clinically important target proteins have been developed and used as therapies to a wide variety of diseases. Monoclonal antibodies are usually produced in non-human species, most often in mice. However, to become successful biotherapeutics, these non-human antibodies have to overcome several significant drawbacks. First, non-humanized antibodies can elicit an immunogenic response when administered to patients, as the human immune system can recognize the non-humanized antibodies as foreign. Creation of chimeric antibodies (non-human variable regions with human constant regions) can partially alleviate these problems. A much more effective strategy is humanization of the antibody. This process involves transferring the antibody CDRs into a proper human antibody framework, while retaining its original affinity and specificity. LakePharma has developed and validated an efficient platform for antibody humanization that combines the advantages of both rational and empirical approaches.
LakePharma has a keen focus on continuous improvement, developing and implementing advanced technologies that have produced the highest standards in quality and efficiency. In 2013, LakePharma published an article in the journal BMC Biotechnology, describing the development and use of the T20 score analyzer to quantify the humanness of the variable region of monoclonal antibodies. (Gao SH, Huang K, Tu H, Adler AS. Monoclonal antibody humanness score and its applications. BMC Biotechnology. 2013;13:55. doi:10.1186/1472-6750-13-55.)Click here to view the full article in BMC Biotechnology.Using LakePharma's analysis tool, the "T20 Score Analyzer," users are able to calculate a monoclonal antibody humanness score from antibody variable region sequences. The T20 Score Analyzer clearly distinguishes human and non-human antibodies with excellent specificity.Click here to access the T20 Score Analyzer.
1. Antibody sequence analysis and homology modeling of mAb 3D structure.
2. Identification of key positions supporting CDR loop structure and VH-VL interface.
3. Design humanized variants (3VH, 3VL).
4. Assess the humanness of humanized variants by T20 humanness score developed at LakePharma. [Sean H Gao, Kexin Huang, Hua Tu, and Adam S Adler. (2013) Monoclonal antibody humanness score and its applications. BMC Biotechnology, 13:55]
5. Construct and express nine humanized antibodies (each combination of three designed heavy chains and three designed light chains) as well as a chimeric version (rodent variable regions, human constant regions) at the 10 mL scale in HEK293 cells (Tuna293™ Process).
6. Assess expression levels and measure antigen binding by direct ELISA (24400 only).
7. Measure affinity by Biacore, Octet, or competitive binding and compare to chimeric (24401 only).
1. Sequences of VL and VH
2. Antigen and assay for measuring binding and affinity
1. Study report, including sequences of the humanized antibodies
2. All remaining purified antibodies