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 (STEMTM), 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 (STEMTM). With Abwiz, you get the affinity, specificity, and functionality your project requires.
Here’s how we do it:
Following custom monoclonal antibody production, we initiate our fully optimized monoclonal antibody humanization workflow.
- Selection of human germline frameworks: grafting the heavy and light chain CDRs of the selected clones to a human framework using bioinformatics analysis. We consider the amino acids tolerated within the human framework to avoid compromising immunogenicity.
- Gene synthesis & recombinant IgG production: Lead candidates are directly cloned into IgG expression vectors and are expressed in HEK293 cells.
- Further humanization & affinity maturation with STEMTM: Mutations are introduced to further humanize the CDRs and panned against the 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 anima studies.
Want more detail? Learn more about our affinity maturation services and STEMTM technology.
Starting with immunization, in less than 10 months, you’ll have a humanized monoclonal antibody—optimized and validated—and all we need is the amino acid sequence of your antibody to get started.
Benefits of Our Antibody Humanization Service
We’ve optimized every step of our process to deliver high-quality humanized mAbs for even the toughest therapeutic targets—and you can get the antibody function and characteristics that you need at maximal cost-effectiveness with our fee-for-service, royalty-free payment options.
Humanize your entire antibody
After CDR grafting to a human framework, our novel STEMTM protocol optimizes affinity and specificity while further humanizing CDR sequences.
Optimal functionality & quality
We validate our humanized mAbs using flow cytometry, ELISA neutralization assays, Octet affinity measurement, and functional assays - always working with the client to deliver high-quality antibodies that work.
Scale up-ready
To optimize expression, we incorporate developability screening into our phage selections and library design, allowing you to confidently scale your lead candidates.
Humanization paired with STEMTM delivers superior mAbs
With Abwiz, you get humanized mAbs with improved affinity and neutralization potency. By combining both our functional antibody CDR database and STEMTM —our 3-stage antibody affinity maturation platform—we consistently deliver humanized mAbs that deliver quality results in R&D workflows.
The number of residues targeted for mutation within each CDR for humanization is constrained to allow functional screening of the entire theoretical diversity while maximizing sampling of the potential sequence landscape.
Customer Success Story
Surrogate virus neutralization test (sVNT) data using Gamma spike trimer and ACE2 proteins.
Humanized antibodies as COVID-19 therapeutics
A humanized anti-SARS-CoV2 neutralizing antibody clone hN2Y was engineered using STEM technology to gain improved neutralization of Gamma variant.
The clone LxC1-G10 showed much higher neutralization of Gamma variant (IC50 2.69 ng/mL, IC80 13.30 ng/mL) compared to the original hN2Y (IC50 220.51 ng/mL, IC80 >1000 ng/mL).
Timeline
| Steps | Deliverables | Time | ||
|---|---|---|---|---|
| Humanization |
•Selection of human germline frameworks •Gene synthesis •Recombinant IgG production •Evaluation |
•IgG ELISA data • ~1mg antibody •Affinity measurement (optional) |
3-4 weeks | |
| Affinity Maturation and Optimization |
•STEMTM (STage Enhanced Maturation) technology •CDRs are further humanized and panned against the target antigen with increasing stringency to obtain higher affinity and specificity •Heat treatment is applied to ensure stable clones for better expression and storage |
•Fab ELISA data •VH/VL sequence data •IgG production and validation 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.
Pricing
Interested in combining discovery work with humanization and affinity maturation services? View the pricing of earlier stages of the antibody development plan here.
FAQ
Yes! The Abwiz Bio team has deep rooted experience developing and optimizing rabbit mAbs. Our expertise paired with our novel humanization, Rabwiz and STEM methods make rabbit antibody development and humanization routine and optimized.
Our proprietary phagemid vector is engineered to optimally express native rabbit Fab fragments that have an extra di-sulfide bridge between the 2 cysteines in the framework 3 and CK1 of light chains (dramatically improving the diversity of obtained clones).
Through systematic sequencing and cloning of candidate clones into a mammalian expression vector and transient transfection in HEK293 cells, we can quickly humanize (within 2 months) rabbit antibodies and further improve affinity and specificity with our unique STEM method.
Traditional methods separate the humanization and affinity maturation processes. Uniquely, our novel STEM 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!
