What You Will Learn

Implementing High Throughput SPR into Antibody Discovery and Engineering Workflows

Abstract: Technological advancements and improvements to the antibody workflows within the Antibody Discovery group have led to generation of hundreds to thousands of clones for an individual campaign. Characterizing these to identify clones of interest has thus created bottlenecks in the discovery workflow process. Implementing Carterra's technology into the discovery workflow has enabled HTP binning and binding kinetics with small amounts of material resulting in faster identification of potential therapeutic antibody candidates.

Networking Break

Developability, Kinetic and Binning Properties of 43 Clinical-stage mAbs

Abstract: This talk will reveal the diversity of binding (kinetics and binning) and developability (stability, self-association, hydrophobicity and polyspecificity) profiles for an important class of antibody drug candidates. There will be an emphasis on the use of high-throughput instrumentation (Biacore 8K, Carterra LSA and Meso Scale Discovery) and techniques (AC-SINS and DSF) for early discovery characterizations.

Lunch

Efficient Kinetic Screening of mAb Target Binding by High Throughput Array SPR

Abstract: Measurement of the kinetics and affinity for mAb binding to its target antigen is key to the identification of therapeutic candidates with desired activity and function. Throughput, resolution and sample consumption have been limiting factors for detailed kinetic characterization early in the mAb discovery process. We will discuss the binding results for a large panel of hybridoma supernatant samples binding to a 35 kDa monomeric antigen on the Carterra LSA array. Within a single day’s run, 384 independent kinetic measurements were completed on an array comprised of 84 unique mAbs, each immobilized at multiple capacities using a capture format binding to an antigen concentration series with >2000-fold range to enable reliable analyses for a wide range of affinities. This Carterra LSA method requires <1 μg per mAb and only 20 μg of the recombinant antigen. When compared to those obtained using the Biacore 8K instrument, high quality kinetic data were obtained with excellent correlation between the results (k_a, k_d, K_D) from both platforms. The Carterra LSA is ~ 5-fold higher in throughput and requires ~10 times less antigen material than the Biacore 8K. Furthermore, several samples that didn’t yield reliable kinetics due to extremely low capture on the Biacore 8K produced results from the LSA analyses potentially due to its continuous flow microfluidics (CFM) technology. The LSA analyses provided reproducible kinetic measurements for supernatant samples tested in replicates with a 4,000-fold affinity range for the target antigen. The combined characteristics of the Carterra LSA array, e.g. quality and reproducible kinetic analysis, high throughput, minimal sample usage (mAb and antigen), capacity to work with unpurified dilute samples, demonstrates its significant potential as a high throughput kinetic screening platform to enhance the early discovery process for therapeutic mAbs.

Evaluation of the Carterra LSA for High Throughput Antibody Affinity Screening

Abstract: Reliable methods are essential for affinity screening of a panel of antibody molecules for a therapeutic antibody generation pipeline. The method should be suitable for screening of the unpurified antibodies from hybridoma supernatants. We used a panel of antibodies present in hybridoma supernatants, binding for a target protein and collected in 96 well plates to evaluate Carterra’s LSA for high throughput affinity screening. We also evaluated the lowest concentration of the antibody required to generate a good kinetic data. The results of this study will be discussed.

Round table discussion with the panel of speakers

Networking Break