Senior Representative, Twist Bioscience

Twist Biopharma, a division of twist Bioscience, combines HT DNA synthesis technology with expertise in antibody engineering to provide end-to-end antibody discovery solutions — from gene synthesis to antibody optimization. The result is a make-test cycle that yields better antibodies against challenging targets from immunization, libraries, and machine learning. Twist Biopharma will continue to optimize and expand its discovery, library synthesis and screening capabilities in partnership with others to further utilize their make-test cycle.

• Mammalian display provides a unique platform to screen new and engineered antibody binders ­­for affinity and developability in the same campaign
• Mammalian display combined with phage display allows selection from large libraries
• Orion’s mammalian display platform is able to select variants with improved biophysical properties and recent developments open for new ways to generate unforeseen sequence variants

At LifeArc we carry out high-throughput antibody screening to discover, characterise and deliver therapeutic monoclonal antibodies. We perform detailed kinetic profiling, epitope specificity, cross reactivity, functional binding and a suite of developability assessment assays to ensure successful downstream development of our lead molecule. We are always looking at ways to innovate our screening platform to ensure we remain at the cutting edge of technology, enabling us to characterise more molecules in a more efficient process. Recently we have purchased the Carterra LSA and here we will share the evolution of our screening platform and how the LSA will enhance our discovery workflow.

Bi- and multispecific antibodies are important molecules for new therapeutics not only for cancer immunotherapy but have also a high potential for other indications ranging from infectious diseases to bleeding disorders. Single domain antibodies are the smallest robust antibody units. The talk will present a comprehensive comparison of formats for the generation of robust bispecific antibodies through fusion of single-domain antibodies on IgG scaffolds in combination with Fab. It comprises a toolbox of complementary methods for in-depth analysis of key features, such as in-solution dual antigen binding, thermal stability, and aggregation propensity, to ensure high bsAb quality. Furthermore, a set of novel in-silico designed humanized single-domain antibody phage display libraries with maximal functional diversity and CDR3 lengths from 10 to 25 aminoacids for generating fusion partners will be presented.

Therapeutic antibody producing Chinese Hamster Ovary (CHO) cell lines have been pushed to their limit, leaving developability and formulation groups to stabilise biologics that were not designed with aggregation in mind. Aura+ is the first instrument designed to characterise antibody stability as early as cell line development (CLD). Aura enables low volume, high throughput subvisible particle imaging, counting, sizing, and identification. It easily handles and analyses the biologically complex cellular and protein samples present in CLD to characterize protein stability at the point of production, following release from CHO cell lines. In this talk, we will show how Aura+ quickly and accurately characterizes secreted antibody stability during CHO CLD in unfiltered cell line samples and using the same antibody labeling protocols established by the innovators to characterise protein titre to rank cell lines according to the physical stability of secreted antibodies using volumes as low as 40 µL per sample. In addition, the Aura+ brings with it the capability to rapidly assess different formulation strategies and, for the first time, distinguish polysorbate degradants from other protein aggregates through the application of a novel fluorescent workflow.

Physics-based free energy perturbation (FEP) calculations provide accurate energetics while allowing conformational flexibility by using explicit solvent molecular dynamics (MD) simulations with a state-of-the-art force field. The accuracy and efficiency of these calculations can be improved through enhanced sampling protocols for mutating residue and nearby waters, effective handling of proline and charged amino acids, and automated parameterization of non-canonical amino acids. FEP+ and our new constant-pH molecular dynamics (CpHMD) implementation can account for protonation and tautomeric state changes, both upon binding/folding and at different pH values. Our approach was recently demonstrated in a real-world collaboration, where it was able to reduce cost and accelerate the development process significantly.

Affibody molecules are small engineered alternative scaffold affinity proteins that can be site-specifically loaded with cytotoxic drugs to create homogenous conjugates with a desired drug-to-carrier ratio. The presentation will explore the targeting of HER2 and HER3 with affibody-based drug conjugates. It will also describe the impact on biodistribution and in vivo cytotoxic efficacy of drug conjugates loaded with auristatin and maytansine-derived payloads.

Developability assessment of monoclonal antibody (mAb) candidates is a critical early process to improve the likelihood of selecting a biologic therapeutic that reaches the clinic. Many biophysical and biochemical attributes are of interest in the assessment process, and with advances in machine learning, it is possible to predict how alterations made to the primary sequence of a mAb impact these traits, in particular stability. Here, the Protein Sciences Department within Biologics Discovery at Merck & Co. Inc used NanoTemper’s Prometheus Panta to collect and rank parameters related to the conformational and colloidal stability of their mAb candidates to enhance their developability profiling.

The goal of biologics drug discovery is to maximize clone diversity to increase the chances of finding and selecting a manufacturable and potent Development Candidate fast. Depending on the method used to capture clones (e.g., hybridoma libraries, single B cell screen, immunized libraries, in vitro translation), different biases and losses of diversity will be evident. Traditional and newer technologies will be discussed. Case studies from biologics drug development, where library design and sources of V regions affect the success of identified leads, will be presented

• A subset of antibodies harboring ultralong CDR-H3 regions can be found in cattle. To investigate the applicability of ultra-long CDHR3 antibody identification using FACS technology, we combined cattle immunization with yeast surface.
• Obtained EGFR antigen-specific antibodies revealed a broad epitope coverage and distinct binding properties compared to canonical reference antibodies. The cattle-derived chimeric antibodies were also potent in inducing NK-mediated ADCC against EGFR-overexpressing tumor cells and most of the antibodies were able to significantly inhibit EGFR downstream signaling. Additionally, we could demonstrate that a minor fraction of CDR-H3 knobs derived from generated antibodies was capable of independently functioning as paratope against EGFR, largely maintaining the binding and functional properties.
• We were also able to engineer ultralong CDR-H3 common light chain bispecific antibodies targeting EGFR on tumor cells as well as NKp30 on NK cells. Biochemical characterization of the bispecifics revealed highly specific binding to the respective antigens as well as simultaneous binding to both targets. Most importantly, engineered cattle-derived bispecific common light chain molecules elicited potent NK cell redirection and consequently tumor cell lysis of EGFR-overexpressing cells as well as robust release of proinflammatory cytokine interferon-γ.