Antibody Humanization for Brain Tumors

With the rapid development of biotechnology, the development of antibodies has progressed from mouse-derived antibodies, chimeric antibodies, to humanized antibodies, and now to fully humanized antibodies. The emergence of humanized antibodies and fully humanized antibodies has brought new hope for clinical applications in brain tumors. These include reducing the adverse effects of traditional brain tumor drug therapy and serving as a brain tumor-specific marker.

We provide humanization service for antibodies in brain tumors

Humanized antibodies are based on chimeric antibodies that further expand the humanized region of the antibody, with humanization rates of up to 80%-90%. Alfa Cytology provides our customers with humanization services for antibodies used in brain tumor research, which allows antibodies to be used in brain tumor research with reduced human allogeneic rejection. In addition to this, we are actively trying to provide fully humanized brain tumor antibodies by using animal gene knockout and insertion and phage display technologies.

• Animal knockout and insertion. We use knockout technology to knock out animal antibody genes, resulting in the deletion of animal antibody genes. Human antibody genes are moved into antibody gene-deficient animals by transgenic or transchromosomal techniques. Full humanization of antibodies for brain tumor research is achieved through the expression of human antibodies in animals.
• Phage display technology. We insert the human antibody variable region gene into the appropriate position of the phage shell protein structural gene, and the human antibody variable region is expressed with the expression of the phage shell protein and, at the same time, displayed on the phage surface with the reassembly of the phage. The fully human antibody is then obtained by display library screening and cellular expression.

General service flow

When designing humanized antibodies, we change key amino acid residues in the human framework residue (FR) region to animal-derived FR to reduce the impact on the complementarity-determining regions (CDR) structural domain.

Screening of humanized antibodies for brain tumors

• Expression levels of mammalian cell protein expression systems (HEK293F or CHO) for transient expression.
• Comparison of humanized antibody binding to chimeric antibody binding based on assays EC50 (by ELISA or FACS) or Kd (by Biacore or Octet).
• Compare humanized antibody activity to chimeric antibody activity based on in vitro bioactivity assays.
• In vitro binding assay based on which humanized antibodies are tested for cross-reactivity with related homologous species.
• Compare humanized antibodies to chimeric antibodies based on biophysical property analysis. This includes identification of the degree of macromolecular aggregation by SEC-HPLC, reductive and non-reductive SDS-PAGE identification, and detection of Tm values by DSC analysis.

Alfa Cytology can humanize mouse-derived brain tumor monoclonal antibodies and other antibodies using CDR substitution technology and computer-aided structural simulation design to ensure >90% humanization. Please contact our staff to learn more about how we can provide our customers with quality brain tumor antibody humanization services.