What You Will Learn

Abstract: Surface plasmon resonance (SPR) is the de facto technique for measuring the binding kinetics and affinities of antibody interactions but has been relegated to a secondary role in discovery due to its limited throughput. Here, we introduce the Carterra LSA, a high throughput SPR platform that is disrupting antibody analytics by enabling screening and characterization to be performed in the same step. The LSA can measure the binding kinetics and affinities of hundreds of interactions in parallel and perform comprehensive epitope binning experiments on up to 384 antibodies per chip. With the additional advantages of minimal sample consumption and industry-leading analysis software, the LSA is streamlining the library-to-leads triage, saving time, cost and resources in generating clinically-ready molecules.

Abstract: Increased funding pressure on drug manufacturers has led to an increased need for SPR machines with higher sensitivity and increased throughput that enable comprehensive antibody characterisation through kinetic screening, epitope mapping and epitope binning. Early adoption of high throughput SPR into antibody discovery platforms leads to increased information about binding and kinetics that can have a profound impact upon identifying the best antibodies with unique epitopes to take forward through HIT, Lead, Lead optimisation and characterisation. Ultimately, this saves both time and money during the drug discovery process. As antibody-antigen affinity can be ‘tuned’ using standard protein engineering techniques, the low protein consumption, ease of assessing multiple parameters such as off-target binding and toxicology differences coupled with the ability to generate meaningful and accurate data quickly makes SPR an choice. Despite the advancement in machinery, SPR can still be viewed as an ‘art’ instead of a ‘science’ due to the lack of exposure to the technique by graduate and postdoctoral students. Ultimately this lack of exposure leads to the basic knowledge about what key steps are involved in designing and optimising a robust and appropriate assay being missing for a large number of SPR users. In this presentation, I will look back at the origins of best practices in SPR and show that when they are applied to assay setup virtually identical data can be generated independently on two leading SPR machines by different users working independently in different laboratories.

Networking Break

Guangwei Yang
Principal Scientist
Boehringer Ingelheim

From Epitope Binning to Function: Case Studies of Antibody Discovery and Characterization at Boehringer Ingelheim

Abstract: Advances in antibody discovery and engineering have enabled the rapid generation of large antibody libraries with therapeutic potential.  These large libraries require high throughput analytical techniques to characterize the interactions between the antibodies and the therapeutic targets. We are establishing new LSA-based processes by incorporating high throughput affinity screening and epitope binning into the early antibody discovery screening process. This new process, combining with functional analysis, enables a robust workflow for lead identification. Results from an ongoing project executing this new workflow is used to demonstrate the correlation between epitope bins and biological function, which in turn allows for more educated decision making for therapeutic antibody development.

Judicael Parisot
Senior Applications Scientist
Carterra

Strategies and Approaches for Detailed Label Free Kinetic Analysis using High Throughput SPR

Abstract: While the epitope is typically the fundamental functional property for determining an antibodies mechanism of action, binding kinetics and affinity is a key parameter for ranking and predicting potency of clones within an epitope class. The detailed kinetic analysis of large numbers of clones has historically been difficult when the antibody samples are limited, was very time consuming, and consumed large quantities of antigen. The Carterra LSA is an ideal tool for high throughput kinetic screening and analysis, and solves these problems with the ability to analyze real time kinetics to 384 mAbs in parallel, from small amounts of crude sample, using a single injection of an antigen concentration series. This presentation will explore common approaches for performing high throughput kinetic screening and approaches to assay optimization. Topics will include strategies for preparing an ideal capture surface, assay format selection, how to create a concentration series which will allow for rate constant determinations over broad kinetic range, and strategies for data analysis and presentation.

Lunch

Judicael Parisot
Senior Applications Scientist
Carterra

Best Practices for Designing and Analyzing Epitope Binning Experiments Using the Carterra LSA and Epitope Software

Abstract: Here, we will describe practical guidelines for designing, executing and analyzing epitope binning assays using Carterra’s LSA and industry-leading Epitope™ software. While the traditional output of a binning assay is a 2D-heat map, this is an over-simplification because antibodies are probing the 3D-topography of an antigenic surface. Heat maps can often appear incomprehensible since there can be significant cross-talk of epitope clusters, but the true complexity of the epitope coverage can be confounded by artefactual complexity introduced by poor assay design or poorly behaved antibodies. Furthermore, epitope competition is not always a binary process of block or not block, and should instead be viewed as a spectrum of behaviors where two antibodies can perturb one another’s binding via steric and allosteric mechanisms, and these behaviors manifest in an assay as asymmetric blockade and displacements. Our Epitope software provides a variety of visualization tools to aid conceptualization of the true epitope diversity, including network plots, dendrograms, and custom communities. We will provide tips and tricks for leveraging the power of our software to groom a data set and remove unnecessary complexity while retaining sufficient nuance to most accurately describe the epitope landscape. 

Networking Hour