Custom Antibody Services
Antibody Humanization Service
Obtain scale up-ready mAbs that are optimized and validated for your specific application with our antibody humanization services. By pairing humanization with our proprietary affinity maturation technology (STEM™), Abwiz delivers the highest-possible quality antibodies needed to accelerate your speed to the clinic.
Overview
As part of our end-to-end mAb development solutions, we offer antibody humanization services to transform your mouse, rabbit, llama, or alpaca antibodies into promising therapeutic lead candidates. Unlike conventional services that only perform humanization of antibodies, our method allows us to simultaneously develop better antibodies by combining humanization with our multi-stage affinity maturation service (STEM™). With Abwiz, you get the affinity, specificity, and functionality your project requires.
Here’s how we do it:
- Selection of human germline frameworks: The heavy and light chain CDRs of your clone are grafted onto a human acceptor germline framework, chosen to minimize immunogenicity and maximize developability of your humanized therapeutic. In silico modeling and bioinformatics ensure that the resulting clone retains the function of the parent mAb while maximizing its humanness.
- Gene synthesis & recombinant IgG production: Lead candidates are cloned and expressed as IgG for functional screening by ELISA, flow cytometry, biolayer interferometry affinity assessment, or by cell-based functional assay as required.
- Further humanization & affinity maturation with STEM™: Back-mutations to reflect natural human immune repertoire amino acid usage are introduced to further humanize the CDRs. The humanized library is panned against target antigen with increasing stringency to obtain higher affinity and specificity or to gain cross-reactivity to the same antigen in other species (e.g. cynomolgus, mouse) for animal studies.
Want more detail? Learn more about our lead optimization services.
In less than 4 weeks, we can have your humanized IgGs validated using a functional assay of your choosing—and all we need is the amino acid sequence of your antibody to get started.
Humanization paired with STEM™ delivers superior mAbs
Uniquely, we can not only humanize your mAb’s framework residues, but we can use our patent-pending lead optimization platform to further humanize your CDRs while simultaneously selecting for clones with improved function.
Competitors only graft your CDRs onto a human framework – this can leave immunogenic regions from the origin species – causing manufacturing and immunogenicity concerns during downstream therapeutic development. STEMTM can not only create humanized mAbs with improved function, but the resulting engineered clones will possess CDRs that reflect more natural human immune sequences.
Timeline
| Steps | Deliverables | Time | ||
|---|---|---|---|---|
| Humanization | • Selection of human germline frameworks • Gene synthesis • Recombinant IgG production • Functional evaluation |
• IgG validation data • ~1mg antibody • Affinity measurement (optional) |
3-4 weeks | |
| Affinity Maturation and Optimization | • STEM™ (STage Enhanced Maturation) technology • CDRs are further humanized and engineered for higher affinity and specificity • Developability filters let you proceed directly to therapeutic mAbs |
• Fab validation data • Full sequence data • IgG production on request |
2-4 months |
Interested in combining discovery work with humanization and affinity maturation services? View the timeline of earlier stages of the antibody development plan here.
Example
As a leader in the antibody discovery field for over a decade, we have extensive experience humanizing antibodies from mouse and rabbit sources. When choosing a humanization strategy, it is important to consider all residues that may impact antigen binding, including the less well understood ‘complementary determining region 4 (CDR4)’ within heavy chain framework region 3. These residues are near the antigen binding site and can either directly interact with the antigen or can be critical to positioning CDRs for proper antigen engagement.
In the below example, we humanized an Ebola neutralizing antibody discovered in macaque. Our initial CDR-grafted humanization design showed much weaker binding to recombinant Ebola spike protein by ELISA. However, when we introduced three residues from the parent CDR4 into our humanized candidate, we regained nearly the full activity of the parent macaque mAb.
We routinely perform antibody humanization within our own therapeutic mAb development pipeline, so we understand the need to not only retain the activity of your parent mAb but also to consider downstream aspects including immunogenicity and developability. These key concerns are always incorporated into our humanization strategy.
FAQ
Yes! The Abwiz Bio team has deep rooted experience developing and optimizing rabbit mAbs. Our expertise paired with our novel humanization, RabwizTM and STEMTM methods make rabbit antibody development and humanization routine and optimized. See here for an example of a broadly-neutralizing COVID-19 therapeutic mAb that we discovered from rabbits and fully humanized and optimized in-house.
Rabbit mAbs possess pM KD affinity and are well-stablished as excellent sources for therapeutic mAb development (Link). Despite possessing exceptionally high diversity within their CDRs, we typically observe that our lead candidate rabbit mAbs are derived from only one or a few germline genes, making humanization relatively straightforward.
Traditional methods separate the humanization and affinity maturation processes. Uniquely, our novel STEMTM affinity maturation approach builds CDR-targeted libraries that are enriched for more ‘human’ amino acids at each position. During affinity maturation, this allows us to not only engineer higher affinity and better function, but also further humanize the antibody sequence for a better therapeutic candidate.
Yes - we optimize developability during every aspect of the campaign. CDR grafting is carefully performed to ensure that the best human framework(s) are chosen, and CDR-targeted libraries are designed to avoid sequence liabilities that could compromise expression or immunogenicity. Finally, our robust developability selection protocol, employing thermoresistance stress during phage panning, ensures that selected clones retain good biophysical characteristics upon IgG cloning and expression in mammalian cells.
Each lead candidate is thoroughly characterized using multiple methods:
- ELISA
- Full VH and VL sequence analysis
- Flow cytometry
- Octet Bio-Layer Interferometry (BLI), on request
- Functional Assays, on request
Our dual-purpose vector allows soluble Fab expression in 96-well format, so we can screen hundreds of lead candidates in high-throughput using crude bacterial supernatant before proceeding to the expensive, time-consuming IgG stage. Even rare clones will be captured!
Get in touch today to discuss your project requirements- we'd love to help.
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