Antibody engineering

Build better binders

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The making of therapeutic antibodies

Antibodies represent one of the most prevalent therapeutic approaches, with a wide range of variations available to optimize function across oncology, neurology, and immunology. The successful transition from development to clinical application of antibodies depends on optimizing antibody stability, target specificity, and the ability to modulate immune responses.

Achieving this requires simultaneous engineering of a candidate’s stability and affinity against its target and other components of the immune system. The study matrix changes through the drug development pipeline, from simple buffers to biofluids and finally to the adjuvant-heavy buffers used in formulation.

The technology used for the assessment, therefore, needs to be robust and flexible for diverse types of interactions, diverse types of samples (purified or non-purified, in buffers or in serum, lysate or feedstock) and have built-in quality control features to ensure a streamlined measurement process across all stages of antibody development.

How can we help?

We built our Fluidity One-M instrument with your challenges in mind. Our proprietary Microfluidic Diffusional Sizing (MDS) technology enables in-solution measurements of antibodies and their targets across all stages of the development, while the smart assistant – Fluidity Insight – with advanced machine learning experiment guidance, will ensure you’re always on the right path.

True size

Measure changes in size to study affinity, antigen conformation and antibody stability. Unlock competition experiments and study ternary complexes.

True environment

Identify high titer clones in cell culture. Comprehensive titer and affinity testing of antibody expression in fermentation media. Assess clinical-stage antibodies in serum.

True insights

Learn how mAbs bind to certain epitopes or how bispecific antibodies engage with the target antigen in solution.

Application Note

Binding of Fc-engineered IgG antibodies to FcRn

Quantifying human IgG-Fc receptor binding using surface-based, kinetic biomolecular interaction analysis has proven difficult due to reagent heterogeneity resulting in complex binding kinetics with multiple KD values. In this application note, we present a straightforward equilibrium binding assay that yields easy to interpret and universally comparable KD values for IgG–Fc receptor
binding affinities.

True size

Discrimination between bidentate and cross-linking binding was readily achieved using Microfluidic Diffusional Sizing. Direct detection of the size of the complex formed between a heteromeric bispecific binder and SARS-CoV-2 spike protein clearly showed that cross-linking was not taking place.  

Read more from the original article.

Figure adapted from Taylor et al., used under Creative Commons 4.0 license.

True environment

Early identification of antibody candidates that are prone to non-specific interactions can avoid significant wasted time and effort later in the development process. There is, however, a paucity of techniques and processes for this critical task. Researchers from the University of Cambridge and Novo Nordisk developed an innovative approach using Microfluidic Diffusional Sizing to challenge antibodies with the non-specificity detecting molecules insulin and a DNA oligomer. Detection of binding to both agents correlated with poor developability profiles.

Read more from the original article.

True insights

Analysis of binding events using surface-based technology can suffer from a range of artefacts. Steric hindrance of binding sites can cause an originally homogeneous sample to show multiple different affinities, a problem that is frequently observed when measuring interactions between antibodies and Fc receptors. Characterizing these interactions in solution with Microfluidic Diffusional Sizing allows robust and easy determination of their true affinities without artefactual interference.

Read more from our Application Note.

Explore related publications

Characterize protein conjugates

Synthetic bioconjugation approaches allow access to structures that are inaccessible via genetic modification methods alone. Taylor et al. achieved site-specific conjugation under mild conditions using a four residue π-clamp motif to enhance the reactivity of their coupling agent. Using this approach they created dimers of designed single-domain antibodies targeting SARS-CoV-2 receptor binding domain. While the individual antibodies had modest affinity, exploiting avidity by generating dimers yielded tight binders.

Taylor, Ross J., et al. “π-Clamp-Mediated Homo- and Heterodimerization of Single-Domain Antibodies via Site-Specific Homobifunctional Conjugation” Journal of the American Chemical Society 144 (2022): 13026-13031

Identify antibody candidates prone to aggregation

Nonspecific interactions of antibodies can impede or even entirely block their development to successful biotherapeutics. Herling et al. have developed a promising method to identify antibodies with low developability that could be applied early in the drug development pipeline by measuring the size change of candidates in the presence of two nonspecific interaction probes. They apply this method to a panel of 12 antibodies of differing properties and developability to validate the approach.

Herling, Therese W., et al. ” Nonspecificity fingerprints for clinical-stage antibodies in solution.” Proceedings of the National Academy of Sciences USA 120 (2023): e2306700120

Fluidic Sciences Ltd