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
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Figure adapted from Taylor et al., used under Creative Commons 4.0 license.
True environment
Read more from the original article.
True insights
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