Recombinant antibody technologies have witnessed remarkable advancements and demonstrated significant clinical success. However, a major challenge associated with the use of murine monoclonal antibodies as a therapeutic is their immunogenicity causing a human anti-mouse antibody (HAMA) response. This response occurs when humans react to the foreign protein, causing complications from immune complex formations and resulting in the antibody treatment being ineffective. To tackle this issue, chimeric antibodies were created using genetic engineering techniques, combining human constant domains with mouse variable domains that recognized the human antigen of interest, thus preserving antibody specificity.1
Antibody humanization was further developed by modifying the mouse antibodies to be more similar to human antibodies by CDR (complementarity determining region) grafting. CDRs of non-human antibodies are grafted to human frameworks of high homology. Typically, some vernier zone residues (murine residues in framework regions) are also retained as they have been shown to affect CDR loop conformation and antibody affinity.2,3 Ultimately, humanized antibodies carry a lower risk of inducing immune responses in humans than mouse or chimeric antibodies.
The humanization process is as follows:
Sequence design and synthesis
Plasmid construction and preparation
Transient expression
Affinity purification
Stringent quality control
Affinity ranking
Biointron offers an antibody humanization service utilizing the CDR grafting and back mutation platform. Our service offers several advantages, including:
Guaranteed affinity (comparable to parental antibody / no cost if guaranteed affinity is not reached)
Fast turnaround time (only 4-5 weeks to obtain humanized antibodies)
Extensive experience and strong expertise (10+ years in humanized antibody design / we have produced 20,000 antibodies)
Mak, T. W., & Saunders, M. E. (2006). Exploiting Antigen–Antibody Interaction. The Immune Response, 147-177. https://doi.org/10.1016/B978-012088451-3.50009-0
Yaghoub Safdari, Safar Farajnia, Mohammad Asgharzadeh & Masoumeh Khalili. (2013). Antibody humanization methods – a review and update. Biotechnology and Genetic Engineering Reviews, 29(2), 175-186. https://doi.org/10.1080/02648725.2013.801235
Harding, F. A., Stickler, M. M., Razo, J., & DuBridge, R. B. (2010). The immunogenicity of humanized and fully human antibodies: Residual immunogenicity resides in the CDR regions. MAbs, 2(3), 256-265. https://doi.org/10.4161/mabs.2.3.11641
The therapeutic efficacy of antibodies is closely related to their ability to recognize and bind specific epitopes on target antigens. Epitopes, or antigenic determinants, are a group of amino acids or other chemical groups that are part of a molecule to which an antibody attaches itself. Epitope characterization can help reveal the mechanism of antibody binding and apply intellectual property (patent) protection for novel antibodies, in addition to designing antibodies with high specificity and minimal cross-reactivity.
Understanding the differences between antibody specificity and selectivity is essential for designing and interpreting antibody-based assays in research for experimental accuracy and data interpretation. Antibody specificity refers to an antibody's ability to recognize and bind to a particular epitope—a unique part of an antigen that elicits an immune response.
Antibody-based assays are essential tools in biomedical research, providing the means to detect, quantify, and visualize specific proteins or antigens within complex biological samples. These assays' efficacy hinges on the antibodies' precise properties. While affinity, avidity, specificity, and selectivity are fundamental to antibody performance, the ultimate impact of these properties is heavily influenced by the experimental context in which the antibody is employed.
Biologics, particularly antibodies, have become indispensable in biomedical research and therapeutic development. Research-use-only (RUO) biologics play a pivotal role in preclinical studies, providing researchers with the necessary tools to explore antibody functions and therapeutic potential in vivo.
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