RDIT^® Antibody Drug De-immunization Services

Background Services Workflow Working with Creative Biolabs FAQ Resources

Background of Immunogenicity in Antibody Drug

Most of the antibody drugs that have been approved or are undergoing clinical trials have immunogenicity to varying degrees. Generally, there are many internal and external factors that may cause or affect the immunogenicity of antibodies. Aggregation, adjuvant like pollutants, the combined use of drugs, co-medication of the patient, and in some cases the release of cytokines are called exogenous factors. The main intrinsic factors that influence or induce the immune response of therapeutic antibodies include specific carbohydrate side chains that exist in variable and constant regions through glycosylation sites. In addition, immunogenicity is also influenced by other post-translational modifications of antibodies, such as glycosylation, deamidation, and oxidation of amino acid side chains.

A large number of studies have shown that the presence of CD4⁺ T helper cell epitopes in injection molecules is one of the factors related to the improvement of the antibody-drug immune response. Therefore, a group of major engineering effects on antibody molecules is aimed at limiting its immunogenicity by reducing its non-human sequence content (chimerism and humanization) or by de-immunization, including the recognition and removal of T cell epitopes.

Fig.1 An example model for de-immunization. (Ducancel, 2012)

RDIT^® Antibody Drug De-immunization Services in Creative Biolabs

Creative Biolabs provides RDIT^® de-immunization service for antibody drugs in combination with our RDIT^® Immunogenicity Assessment Platform. It adopts a novel, innovative and classic methods to reduce the immunogenicity of candidate drugs by combining immunology and molecular biology technology. With rich experience and deep professional knowledge, we will provide great help for your potential drug candidates to enter clinical trials. Alternative technologies include:

RDIT^® Antibody Sequencing	RDIT^® Antibody Engineering
RDIT^® Antibody Expression	RDIT^® Antibody Humanization

The Workflow of our De-immunization Service

Our services have been widely used to remove T cell epitopes from biotherapy drugs while maintaining their therapeutic efficacy. The following is a brief description of the workflow of our de-immunization service for antibody drug candidates.

Locate the T cell epitopes by our RDIT^® immunogenicity assessment service.
Construct a 3D model of the Fv domain of the antibody. The model will help to study the location of predicted T cell epitopes and to evaluate the effects of substitution on the stability and binding affinity of the antibody. The 3D structure is modeled by homology to the most related structure.
Make multiple sequence alignments of human and murine germline as a source of possible substitutions for T cell epitopes.
Replace the T cell epitopes with the closest human germline sequence according to the multiple sequence alignments. If the possibilities of human germline are exhausted, continue with murine germline and then ad-hoc substitutions. All substitutions will be tested by the method in the next step.
Test the compatibility of the substitutions with the 3D structure of the antibody. Check the affinity of the substitutions with human leukocyte antigen (HLA) allotypes. The overlapping peptides should not be transformed into T cell epitopes. The substitutions should avoid inducing strain in the antibody scaffold.
If all tests are passed, accept the substitution. Otherwise, new substitutions are evaluated and selected.

Working with Creative Biolabs

Creative Biolabs tailor services to each project to ensure that goals are met or exceeded. Experienced project teams are assigned to each study to focus on the progress of the project in order to achieve results in the shortest time. Please contact us for more information, a quote or to schedule a conference call.

Reference

Ducancel, F., Muller, B. H. Molecular engineering of antibodies for therapeutic and diagnostic purposes. Taylor & Francis. 2012, 4(4): 445-457.

FAQ

What is antibody drug de-immunization?
Antibody drug de-immunization is a process aimed at modifying monoclonal antibodies (mAbs) to reduce their immunogenicity without affecting their therapeutic efficacy. This involves altering parts of the antibody that are recognized by the immune system to prevent an immune response that could reduce the drug's effectiveness or cause adverse reactions.
Why is de-immunization important for antibody therapies?
De-immunization helps prevent the development of anti-drug antibodies (ADAs) that can neutralize the therapeutic effects or lead to infusion reactions. Reducing immunogenicity extends the drug's effective lifespan in the body and improves overall treatment safety and efficacy.
How is de-immunization of antibodies achieved?
De-immunization is typically achieved through techniques like altering the amino acid sequence to remove T-cell epitopes, humanization of murine antibodies, or glycoengineering to modify glycosylation patterns that affect immunogenicity. These modifications help to make the antibody less detectable or reactive to the patient's immune system.
What are T-cell epitopes, and how are they related to antibody de-immunization?
T-cell epitopes are peptide fragments of proteins that are presented by MHC molecules to T-cells, triggering an immune response. In antibody de-immunization, segments of the antibody that contain T-cell epitopes are modified to avoid activation of T-cells, thereby reducing the antibody's immunogenic potential.
Can de-immunization affect the functionality of antibody drugs?
There is a potential risk that de-immunization could affect the functionality of antibody drugs. Alterations made to reduce immunogenicity need to be carefully balanced to ensure they do not impact the antibody's binding affinity, specificity, or overall therapeutic action.
What techniques are used to identify immunogenic regions in antibody drugs?
Techniques such as epitope mapping, computational modeling, and immunoassays are used to identify immunogenic regions in antibody drugs. These methods help pinpoint specific parts of the antibody molecule that are likely to elicit an immune response, guiding targeted modifications for de-immunization.
How does humanization contribute to antibody de-immunization?
Humanization involves replacing mouse-derived antibody sequences with human sequences while preserving the antigen-binding sites. This process significantly reduces the immunogenicity of chimeric antibodies used in therapeutic applications, as human proteins are naturally less likely to be recognized as foreign by the patient's immune system.
What is the role of glycoengineering in antibody de-immunization?
Glycoengineering involves modifying the glycosylation patterns on antibodies to enhance their safety and efficacy. Altering glycan structures can reduce immunogenicity by masking epitopes and modifying the antibody's interaction with immune cells, thereby enhancing its therapeutic profile.
Are there any successful examples of de-immunized antibody drugs?
Several de-immunized antibody drugs have successfully reached the market. These antibody drugs have been modified to reduce immunogenicity through humanization and glycoengineering. These drugs show improved clinical outcomes with reduced incidence of adverse immune reactions, demonstrating the effectiveness of de-immunization strategies.