Antibody Humanization Service

Humanization is a critical step in the development pathway of all therapeutic antibodies derived from non-human sources, such as hybridomas. Absolute Antibody’s Prometheus™ antibody humanization service delivers a panel of humanized variants from your clone, guaranteeing to produce a variant with comparable activity to the parent antibody. Our service focuses on enhancing antibody manufacturability, leading to increased expression, lower aggregation, and long-term stability and solubility.

Humanized antibody variant expression yields relative to the original chimeric antibody, shown in black. Download our research poster describing our antibody humanization approach to learn more.

Our Humanization Approach

Antibody humanization involves the transfer, or grafting, of critical non-human amino acids onto a human antibody framework. This primarily includes the grafting of amino acids in the complementarity-determining regions (CDRs), but also other framework amino acids critical for the VH:VL interface and CDR orientation. Humanization is a balance between introducing as much human content as possible to reduce the risk of immunogenicity, while retaining enough non-human content to maintain the original binding activity of the parent antibody.

Another key consideration is the developability of the antibody, which is a combination of immunogenicity and manufacturability. Despite the growing trend to de-immunize antibodies, clinical data give it no credibility. However, manufacturability – which encompasses a range of properties such as expression titer, aggregation, stability and solubility – can be determined empirically. At Absolute Antibody, our Prometheus™ humanization service was developed specifically to deliver antibodies with increased manufacturability.

Download our research poster to learn more about the development of our humanization approach, which was informed by analyzing manufacturability data for more than 100 therapeutic antibodies to identify preferential manufacturing properties. The poster also describes an antibody humanization case study, in which we created 25 different humanized variants with increased titers compared to the original chimeric antibody. The three winning candidates displayed no detectable aggregation, an approximately two-fold increase in activity, and a minimum 16-fold increase in titer compared to the chimeric antibody.

State University of New York researcher, describing our services. Read his full antibody humanization case study here.

Advantages of Our Prometheus™ Humanization Service

The Absolute Antibody team has deep antibody humanization expertise, with some of our founders and scientific advisors even humanizing Campath-1H, the first humanized antibody to be used in the clinic. We offer a royalty-free humanization service with a guarantee of success – if we fail to deliver a humanized antibody with comparable activity to the parent antibody, you will not be charged for the service.

Our service is uniquely focused on delivering humanized antibodies with strong manufacturability, enabling the optimized large-scale production required for clinical use. We guarantee to produce a minimum of 12 humanized variants, and we have completed more than 60 successful humanization projects to date, for customers ranging from small academic laboratories to leading pharmaceutical companies.

To learn more about our process and the benefits to our approach, check out this customer case study, which describes how we helped a State University of New York researcher humanize an antibody with the potential to treat glioblastoma.

Model for antibody-antigen interface

Our Humanization Deliverables

If required, antibody sequences can be determined with our hybridoma sequencing or de novo protein sequencing services. Alternatively, customers can come directly to us with a sequence.

An in silico model of the parental variable domains is generated to enable structure-guided humanization. Sequences are aligned to a panel of human germline sequences selected for preferential manufacturability properties, and the non-human amino acids are grafted onto the human sequence using our proprietary Prometheus™ humanization algorithm. A minimum of 12 humanized variants are created.

We offer four levels of service, ranging from in silico humanization only to a full expression, purification and analytics package, as shown in the table below. Contact us to discuss your project today.

HUMANIZATIONBronzeSilver Gold Platinum
Generation of structural model
In silico manufacturability optimization
In silico humanization
Sequence liability analysis
Expression/purification of parent as human chimeric
Expression/purification of up to 16 humanized variants
Determination of expression yields
Measurement of aggregation levels
Antibodies sent to client for activity testing
ELISA activity screen
Full kinetic analysis to determine KD by Biacore
Full report
Service timeline1 week8 weeks9 weeks 12 weeks