Antibody Maturation Service
Overview Workflow Publication Why Choose Us FAQs Customer Review Related Services Contact Us
Overview
Antibody is an indispensable tool for basic research, clinical diagnosis, and therapeutics. However, sometimes an antibody with the most favorable specificity lacks sufficient affinity for a desired application. Affinity maturation is a process in which antibodies gain improved affinity, avidity, and antipathogenic activity. It is a result of somatic hypermutation (SHM) of immunoglobulin genes in Tfh cell (T follicular helper cell)-activated B cells, which combines with the selection for antigen binding. This iterative process occurs in germinal centers and proceeds for weeks after infection or vaccination. The resulting antibodies can be highly mutated with increased affinity to antigens by an order of magnitude.
In Creative Biolabs, we conduct antibody optimization and antibody affinity maturation service that combines with molecular evolution and high-throughput phage library construction and screening to improve antibody potency and to save the cost of antibody production.
Antibody Maturation by Phage Display at Creative Biolabs
The in vitro affinity maturation has been used to optimize antibodies, antibody fragments or peptide molecules. Here, Creative Biolabs hopes to introduce Antibody Affinity Maturation Service to improve the target binding affinity of antibodies to antigens, based on principles of mutation and selection. In addition, we also offer HIAF-ProNano™ Platform to increase the affinity of single domain antibodies by engagement of protein nanocages without changing physicochemical properties of single domain antibodies. A standard maturation process is performed through the following steps.
Fig.1 An optimized process of affinity maturation in Creative Biolabs.
Phage display is the most commonly applied molecular technology in display and affinity maturation for a wide range of proteins. This method enhances the affinity of recombinant antibody fragments by virtue of random mutagenesis and phage display under stringent conditions. Random mutations are inserted by performing radiation, chemical mutagens, or several rounds of error-prone PCR. After construction of a mutated antibody gene library, affinity selection is started by panning with washing solutions optimized for off-rate-dependent selection. As a result, antibody affinity can be improved by at least 10 folds after each round of antibody affinity maturation through phage display.
Except that, Creative Biolabs can take advantage of computer hardware and modeling algorithms to complete a rational design of antibody structures and functions. With the help of computational methods, sampling antibody conformations and scoring designed antibody variants are convenient in antibody affinity maturation process.
Other Molecular Display Technologies
➢ Bacterial display – The bacterial display presents thousands of copies of displayed protein on the cell surface. The most important advance made by cell-based bacterial display is quantitative screening and high enrichment ratios of proteins bound to fluorescently labeled ligands using fluorescence-activated cell sorting. In Creative Biolabs, our bacterial display retains many helpful features of phage display, including the ease of manipulation, high transformation efficiency, and familiarity of handling E. coli. Aided by this system, clients can obtain affinity maturation (at least 5 folds, even up to 100 folds), expression level and thermostable candidates.
➢ Yeast display – Yeast display can overcome limitations of bacterial hosts requiring of phage and bacterial display, since these methods potentially contain variants due to mismatched human/bacterial codon bias. In Creative Biolabs, yeast display benefits from the advantage of FACS-based selections, yet has the characteristic of eukaryotic protein folding pathways and codon usage greatly similar to mammalian cells. Thus, it may be the most ideal system for the surface display of mammalian cell-surface and secreted proteins, particularly antibodies.
➢ Ribosome and mRNA display – In Creative Biolabs, we conduct ribosome and mRNA display that both apply an in vitro transcription/translation (IVTT) system circumventing growth and transformation of cells. IVTT-based techniques promisingly create much larger libraries (>1012) than cell-based systems and potentially avoid expression bias. Notably, IVTT methods have a potential for affinity maturation via recursive mutation, in which candidates are further mutated after each round of selection.
Publication
This study explores how the immune system produces antibodies with enhanced affinity for their targets. By leveraging next-generation sequencing (NGS), researchers can now reconstruct antibody clonal lineages, revealing the structural mechanisms behind affinity-enhancing mutations. The authors further analyze crystal structures of antibodies bound to their antigens, providing atomic-level insights into how these molecules evolve to effectively neutralize pathogens such as HIV-1.
Key findings demonstrate that increased antibody affinity arises from multiple factors, including:
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Improved shape complementarity between antibody and antigen.
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Greater buried surface area upon binding.
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Formation of new intermolecular interactions.
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Preorganization or rigidification of the antigen-binding site.
These structural refinements collectively optimize antibody-antigen interactions, offering valuable insights for therapeutic antibody design.
Fig.2 Affinity maturation through increased antigen interactions.1
Why Choose Us?
With decades of expertise and a proven track record, Creative Biolabs delivers cutting-edge antibody discovery and engineering solutions. We combine unmatched scientific rigor, innovative technologies, and a client-centric approach to drive success. Our commitment to quality is demonstrated by published data, showcasing significant affinity enhancements while maintaining antibody stability, ensuring robust, high-performance results.
Highlights:
✔ 10-100 fold affinity improvement;
✔ Remarkable homogeneity and biocompatibility;
✔ Making further affinity maturation by directed evolution or computational design;
✔ Multiple tools (ELISA, DSC, SEC-HPLC, etc.) for antibody characterization;
✔ Extensive experience and guaranteed services.
For more information or quotes on our antibody maturation services, please contact us.
FAQs
Q1: What is the minimum affinity improvement I can expect?
A1: While the final affinity improvement depends on the starting antibody and the target, our service is designed to achieve a 10- to 100-fold increase in binding affinity per cycle. We recommend a consultation to discuss the specific potential for your project.
Q2: Can you perform antibody maturation on an antibody from a species other than human?
A2: Yes, our platform is flexible and can be applied to antibodies from various species, including mouse, rabbit, camel, and llama. We can also handle different antibody formats, such as scFvs and Fabs, ensuring we can meet a wide range of project needs.
Q3: How do you address potential developability issues like stability and aggregation during the maturation process?
A3: We incorporate developability screening into our workflow from the start. This allows us to screen for improved stability, solubility, and other critical parameters early in the selection process, ensuring the final antibody is not only high-affinity but also a robust and developable molecule.
Customer Review
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Enhanced Assay Sensitivity
Using Creative Biolabs' antibody maturation service in our research has significantly improved the detection limits of our ELISA assay. The enhanced affinity of our capture antibody allowed us to achieve a 10-fold increase in sensitivity, which was a critical breakthrough for our project. - Dr. Aa Sh
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Solving Developability Issues
The team's detailed characterization of the mutated clones was invaluable. They not only improved the affinity of our scFv fragment but also identified a variant with improved solubility, which solved a major developability issue we had been facing. This level of comprehensive analysis is something you don't always get. - Dr. Je Pr
Related Services
To further support your research and development, we recommend exploring our complementary services, which can be seamlessly integrated with your antibody maturation project:
TCR Generation and Optimization
Creative Biolabs offers a comprehensive, one-stop service for TCR engineering and T cell development, from biomarker identification and optimization to preclinical in vivo testing, with tailored strategies for your needs.
Learn More →
Humanized Models
Creative Biolabs has developed a wide range of humanized mouse models, which harbor human immune cells. These models, alongside CDX and PDX models, are powerful tools for studying potential cancer immunotherapies and provide off-the-shelf inventories for preclinical testing.
Learn More →
Contact Us
Creative Biolabs offers an unparalleled combination of expertise, technology, and a client-focused approach to drive your projects forward. Our antibody maturation service is a proven solution for optimizing your lead candidates and accelerating your path to discovery and development.
Contact Our Team for More Information and to Discuss Your Project
Reference
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Mishra, A. K., and R. A. Mariuzza. "Insights into the Structural Basis of Antibody Affinity Maturation from Next-Generation Sequencing." Front Immunol 9 (2018): 117. Distributed under Open Access license CC BY 4.0, without modification. DOI: https://doi.org/10.3389/fimmu.2018.00117
