The following presentation focuses on key factors to consider to increase both the quantity and quality of the recombinant protein of interest. These include the following: system/organism utilized to generate the recombinant protein, ability to enhance the properties of the subject protein via nucleic or amino acid sequence changes, and purification process utilized, such as standard vs. affinity chromatography methods.

Large-scale transient antibody expression is essential for the progression of UCB’s therapeutic pipeline. For over 10 years, we have routinely used our proprietary cell line CHO SXE for large-scale transient expression of antibodies. Recently, we engineered these cells to be GS knockouts which enabled the rapid generation of stable pools using a transposase system. From these pools we obtain higher yields, with less heterogeneity.

Cell substrate manufacturing changes can generate challenging regulatory questions because of the potential to impact therapeutic protein structure, potency, and/or impurity profiles, which in turn may or may not impact pharmacokinetics, pharmacodynamics, clinical safety, and/or efficacy. This presentation will summarize current knowledge on the impacts of cell substrate on therapeutic protein product quality attributes and present process and product quality data from cell lines expressing non-originator NISTmAb.

Site Specific Integration (SSI) expression systems offer robust means of generating highly productive and stable cell lines for standard mAbs. However, the SSI system is not immune to genetic rearrangements and complex multi-specific modalities can prove challenging for the SSI expression systems. Here we present two SSI cell lines which were characterized to contain off-target genetic events. These events were found to have no impact on the phenotypic stability nor product quality, and both cell lines were deemed to be commercially viable.

• What challenges have we faced? 
• What types of improvements are needed in expression hosts? 
• Strategies for maintaining healthy cultures?
• What might address the future and what is needed?

Join a group (topic of your choice) to share and experience and hear what others have learned:

5:30 pm Think Tank Report-Outs: Listen and Learn

During the Think Tank Table discussions, we shared our experiences and working solutions for end-to-end protein production workflows. Now as a collective community, let’s hear from the table facilitators as they share key discussion points and strategies, and provide a wrap-up of their table’s discussion. What can we take away and apply?

• Determine what your end-product could look like
• Consider the full pipeline of your candidate as early as possible
• Select suitable expression systems early on
• Use regulatory acceptable hosts as soon as possible
  

Reduction of interchain disulfide bonds in monoclonal antibodies (mAbs) is one of the critical issues in large-scale mAb production using Chinese hamster ovary (CHO) cells. mAb reduction susceptibility has been reported to be cell line-dependent, but there are no efficient methods for screening reduction-unsusceptible cell lines. Here we present a novel screening method with Ambr15 for the susceptibility, which allows 48 different cell lines to be evaluated simultaneously.

High-throughput (HTP) workflows are critical elements of large-scale drug discovery campaigns, especially during screening and lead identification. By incorporating automation and informatics tools, HTP workflows can be built to effectively manage tasks, optimize resource usage, ingest large volumes of data, and improve productivity. By capturing and leveraging data from automated processes effectively, we aim to enhance decision-making to positively impact the discovery and development of therapeutics for patients.

We have built automated custom expression and purification systems, allowing accelerated protein production of antibodies and target proteins at microgram-to-gram production scales. These semi-automated workflows allow flexible support of early drug discovery pipelines from earliest production to upscaling of lead panels, providing material for HTP characterization, in vitro and in vivo experiments. Production processes will be exemplified using examples of rapid and scalable support of example projects.

We report the development of a semi-automated platform to express panels of antibodies in high-throughput at mid-scale for developability and functional assays whilst minimizing hands-on lab work. Our platform enables parallel production of 100 mg of material for 96 clones, keeping the discovery funnel wide for longer. We have created digital workflows for sample and data-tracking, enabling data reuse and driving refinement of AI/ML models.

Here, we present a multi-component experimental pipeline designed for efficiency and adaptability within the Generative Unconstrained Intelligent Drug Engineering (GUIDE) program. This semi-automated laboratory integrates processes from molecular cloning to CHO-derived antibody affinity and characterization assessments in high-throughput fashion, emphasizing scalability and speed. Through the generation of comprehensive experimental data, our approach empowers computational models for antibody engineering, marking a significant leap forward in the efficiency of high-throughput antibody development.