Cambridge Healthech Institute’s 14th Annual

Cell Line Engineering and Development

Rapid, Nimble, and Flexible Protein Expression Engineering Tools

January 17 - 18, 2022 ALL TIMES PST

How best to express? The complexity of biotherapeutics and the increasing demands of “faster, better, more economical” resonate with recombinant protein expression and production researchers. To meet these goals, protein expression scientists are exploring new engineering tools to make biology easier to “engineer.” However, many variables still must be considered during this process, including verification and sequence analysis of the gene or protein of interest, codon optimization, vector construction and clone/host selection – a time-consuming and expensive process. Ultimately, these tools must be weighed against traditional expression and production strategies to achieve the desired quantity and quality.

Sunday, January 16

4:00 pm Conference Registration Open (Sapphire West Foyer)

Monday, January 17

7:00 am Registration and Morning Coffee (Sapphire West Foyer)


Session Room: Sapphire 410

9:00 am Organizer's Welcome Remarks

Mary Ann Brown, Executive Director, Conferences, Cambridge Healthtech Institute

9:05 am

Chairperson's Opening Remarks

John Dresios, PhD, Executive Director, Biological Innovations Center; Leidos Technical Fellow & Solutions Architect, Applied Science Division, Leidos Innovations Center, Leidos Inc.
Matthew P. DeLisa, PhD, William L. Lewis Professor of Engineering, Chemical & Biomolecular Engineering, Cornell University

There remains an urgent need for new tools that can overexpress structurally uniform glycans and glycoconjugates. To address this technology gap, cell-free synthetic glycobiology has emerged as a simplified and highly modular framework to investigate, prototype, and engineer pathways for glycan biosynthesis and biomolecule glycosylation outside the confines of living cells. This talk will describe how we are leveraging cell-free platforms to produce human therapeutic glycoproteins and conjugate vaccines.

9:40 am

Building Protein Networks in Artificial Bacteria for Biomedical Applications

Cheemeng Tan, PhD, Chancellor’s Fellow; Associate Professor, Department of Biomedical Engineering, University of California, Davis

It is challenging to create artificial cellular systems that rival the dynamic chemical-biological response and resilience of natural cells. The artificial cellular systems are hybrid material-bacteria systems with superior applications in drug discovery, disease treatment, and basic biological study. Dr. Tan will present his work on controlling the information processing of artificial cellular systems. The work has broad impact on biotechnology applications and disease treatment.

10:10 am

Engineered Regulon to Enable Autonomous Azide Ion Biosensing, Recombinant Protein Production, and in vivo Glycoengineering

Shishir P. S. Chundawat, PhD, Associate Professor, Department of Chemical and Biochemical Engineering, Rutgers University

The biological importance of carbohydrates (or glycans) can be better understood and exploited by creating a robust toolkit. Here, we highlight recent advances in glycoengineered microbial strains/plasmids and enzymes development for in vivo conversion of azido sugars to designer glycans (e.g., Human Milk Oligosaccharides). We discuss how such a toolkit can be used for efficient recombinant protein production as well as directed evolution of carbohydrate-active enzymes (e.g., glycosynthases).

Reference: Bandi et al. 2021, ACS Synthetic Biology, 10:4, 682–689.

10:40 am Networking Coffee Break (Sapphire West Foyer)
11:00 am

Assessing Optimal: Inequalities in Codon Optimization Algorithms

Jeffrey Li, Graduate Researcher, Laboratory of Dr. Rachel Green, Johns Hopkins School of Medicine

Non-native synthetic DNA sequences have become a common resource in recombinant protein work due to the low cost of DNA synthesis. Many of these DNAs are “optimized” through commercial tools despite there being little guidance for what defines an optimal sequence. We surveyed nine optimization algorithms and found significant variability between their optimized DNAs, leading us to propose a set of best practices when designing synthetic sequences.

11:30 am

Synthetic Biology Strategies for Robust Production of Active Proteins

John Dresios, PhD, Executive Director, Biological Innovations Center; Leidos Technical Fellow & Solutions Architect, Applied Science Division, Leidos Innovations Center, Leidos Inc.

Production of high-value protein targets often faces significant challenges due to their poor and variable levels of expression and active folding, as well as the long turnaround times required for their end-to-end manufacturing and testing. This presentation describes strategies for robust production of active proteins that are applicable to a wide range of DNA and mRNA expression formats in the context of cell-free, cell-based and gene therapy platforms.

Ademola Kassim, Product Specialist, Cell Line Development, Cell Line, Media and Testing Solutions, Sartorius

When it comes to the commercialization of biologics, effective, high-quality cell line development (CLD) is essential for success. The key to minimizing risk and optimizing outcome is having a reliable partner that offers experience, expertise, and a comprehensive portfolio of products and services. This presentation will highlight the key challenges and points to consider during biopharmaceutical CLD and the impact early decision-making can have at later stages of the development process.

12:30 pm Session Break
12:40 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:10 pm Session Break


Session Room Change: Sapphire 400

1:45 pm

Chairperson's Remarks

Bjørn Voldborg, MSc, Head, National Biologics Facility, DTU Bioengineering, Technical University of Denmark
1:50 pm

Subcellular Proteomics Unveils New Regulatory Mechanisms Controlling mAb Expression along the Secretory Pathway of CHO Cells

Saumel Pérez Rodriguez, Universidad Nacional Autónoma de México

Genetic engineering of proteins from the classical secretion pathway (CSP) is an alternative for obtaining CHO cell clones with enhanced productivity. However, few CSP targets have been detected from homogenates in previous differential proteomic studies. Hence, we used subcellular proteomics to intensify the identification of CSP proteins associated with cell productivity. Differentially expressed proteins (DEP) from this study participate in protein synthesis and translocation, autophagy, proteasomal degradation, calcium regulation, vesicular transport, ER stress and UPR. About 80% of DEP had not been associated with productivity and their further modulation could have a positive impact on bioprocesses. Supported by IN210419.

2:20 pm

Host and Product-Specific Determinants of Recombinant Protein Yield in CHO

Helen Masson, Research Scientist, Nathan Lewis Laboratory, Pediatrics and Bioengineering, University of California, San Diego

Decades of cell line development and media optimization of CHO cell lines has led to notable improvements in recombinant protein (rProtein) production yield.  However, the expression of some rProteins results in little to no yield and remains a challenge.  Our lab has taken a systems biology approach, including machine learning and network analyses, to elucidate host and protein-specific properties that act as determinants of rProteins yield in CHO.

2:50 pm Find Your Table and Meet the BuzZ Sessions Moderator
3:00 pm BuzZ Sessions with Refreshments (Sapphire Foyer)

PepTalk BuzZ Sessions are focused, stimulating discussions in which delegates discuss important and interesting topics related to upstream protein expression and production through downstream scale-up and manufacturing. This is a moderated discussion with brainstorming and interactive problem-solving between scientists from diverse areas who share a common interest in the discussion topic. Continue to check the event website for detailed discussion topics and moderators.

BuzZ Table 7: Common Issues with Transient Protein Production

Richard Altman, Field Application Scientist, Life Science Solutions, Thermo Fisher Scientific
Henry C. Chiou, PhD, Director, Cell Biology, Life Science Solutions, Thermo Fisher Scientific
  • What are the current challenges to transient protein production?
  • How has the COVID-19 pandemic affected your workflow and productivity?
  • How do we optimize the whole protein expression workflow process?
  • How can we maintain volumetric yields while scaling transient expression up or down?
  • What cell line(s) should we use and when?
  • What parameters can impact the quality or physical attributes of transiently produced proteins?​​


4:00 pm

Apoptosis-Resistant CHO Cell Lines Significantly Improve Culture Viability and Titer in Intensified Fed-Batch Culture Process

Shahram Misaghi, PhD, Principal Scientist, Cell Culture and Bioprocess Operations (CCBO), Genentech, Inc.

Process intensification strategies in CHO production cultures can potentially increase productivity, lower cost of goods, and improve facility utilization. However, process intensification often triggers apoptotic cell death in the later phases of intensified production process. Here we show that apoptosis-resistant CHO cell lines counteract this undesired outcome, resulting in not only better viability but also enabling extended productivity that significantly improve volumetric productivity without affecting product quality.

4:30 pm

Novel CHO Host for Improved Recombinant Protein Production

Lina Chakrabarti, PhD, Senior Manager, R&D, AstraZeneca

With the aim of increasing protein productivity, we generate a novel CHO host with favorable biomanufacturing phenotypes and improved functionality. Producer pools and clones generated from the new host outperformed the standard host by displaying (1) improvement in productivity, (2) reduced product aggregation, (3) enhanced cell viability, (4) low lactate production and (5) improved cell cloning efficiency. Proteomics and western blot analysis demonstrate that the new host acquired multifaceted protection against mitochondrial dysfunction and ER stress.

5:00 pm

Prediction of Amino Acid Consumption in Chinese Hamster Ovary Cell Fed-Batch Cultures by Coupling a Genome-Scale Metabolic Network Model with Machine Learning

Wei Wei, PhD, Principal Scientist, Cell Line Development, Biotherapeutics Pharmaceutical Sciences, Pfizer Inc.

The control of nutrient availability is critical to large-scale manufacturing of biotherapeutics. Genome-scale metabolic modeling offers a promising approach for in silico monitoring and predicting the consumption of proteinogenic amino acids, which is critical for bioprocess control. However, the prediction accuracy is challenged by the discrepancy between the model assumption and the biological variances in CHO cultures. We demonstrate such challenge can be addressed by integrating a CHO-specific metabolic network model with machine learning to achieve accurate prediction throughout the fed-batch process.

5:30 pm

Digital Twins for Improved Bioprocess Operation: Opportunities and Roadblocks

Krist V. Gernaey, PhD, Professor, Chemical & Biochemical Engineering, Danish Technical University

Industry 4.0 can potentially revolutionize biomanufacturing. Digital twins, virtual copies of a process interacting with the process, will play an important role in this transformation. This talk has focus on understanding the needs and challenges faced by the biomanufacturing industry when dealing with this digitalized paradigm. Two major building blocks of a digital twin, data and models, are highlighted. Data characteristics and collection strategies are examined, and new methods and tools for data processing are highlighted. Furthermore, different modelling approaches are presented in view of their use in a digital twin. Potential roadblocks for digital twin implementation are discussed.

6:00 pm Welcome Reception in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
7:30 pm Close of Day

Tuesday, January 18

8:30 am Registration and Morning Coffee (Sapphire West Foyer)


Session Room: Sapphire 410


Cell Engineering Efficiency and Quality

Zhimei Du, PhD, Vice President, Process Sciences, Atara Biotherapeutics

The efficiency of generating bi-allelic gene-knockout cell lines using conventional protocols is very low. This significantly affects clone screening efficiency and reduces the chance of identifying robust knockout host cell lines. In this study, we improve the genome editing process resulting in improved GS-knockout efficiency of up to 20 folds. Furthermore, we developed an integrated end-to-end process yields robust host cells with desired growth and recombinant protein expression characteristics.

9:35 am

CHO Cell Engineering to Improve Therapeutic Antibody Production

Cai Guo, PhD, Scientist, Expression and Cell Engineering, Amgen, Inc.

Bispecific antibodies have become increasingly of interest for therapeutic applications. These antibodies can be difficult to express (DTE) with much lower productivity than classical monoclonal antibodies. Besides protein engineering, vector and bioprocess optimization, CHO host engineering shows potential to enhance DTE protein productivity. Here, we analyzed the transcriptome of cells expressing various DTE proteins and identified genes that were differentially transcribed in bispecific antibody expressing cells. The effects of these genes on productivity were subsequently evaluated by gene overexpression or gene knock out/down. Our results indicate that proper host cell engineering is one solution to improve recombinant therapeutic protein production.

10:05 am Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
Bjørn Voldborg, MSc, Head, National Biologics Facility, DTU Bioengineering, Technical University of Denmark

Based on engineered CHO cell lines generated over the past 9 years, we offer cell lines for quick and robust production of recombinant proteins of high quality and controlled glycosylation. The cell lines can now be accessed by industrial and academic partners through our newly established National Biologics Facility, where we offer establishment of robust production cell lines, protein production, cell line development and cell line engineering.

11:30 am

Improvements to the Baculovirus Insect Cell Expression System to Support Structural Biology and Drug Discovery

Dominic Esposito, PhD, Director, Protein Sciences, Frederick National Laboratory

Insect cell systems have become a major tool in the toolbox for the production of pharmaceutically relevant protein targets. In our laboratory, this system has been vital for production of multiprotein complexes for structural biology and drug discovery research. We have developed a number of process and technology improvements which permit increased protein yield, protein quality, virus stability, and efficiency of protein complex formation. We will discuss in detail the enhancements to the system and how they can be applied to high-level production of other clinically relevant proteins.

Oren Beske, PhD, Amalgamator of Business and Biology, ATUM

Launched only a few years ago, the Leap-In Transposase platform has rapidly become an industry standard technology for the generation of CHO cells for the manufacturing of antibodies and other biologics.  This presentation will highlight achievements and case studies of the platform including high titer mAb manufacturing, rapid anti-COVID responses, and some novel, next generation, applications.  

12:30 pm Session Break
Mingchao Kang, PhD, Group Leader, Platform Technology, Ambrx Biopharma

Ambrx’s EuCODETM platform utilizes engineered CHO cells with an expanded genetic code to produce engineered precision biologics. Quick material delivery with high productivity is critical for rapid project development. MaxCyte’s STX electroporation system enables systemic optimization of transient expression conditions and a facile scale-up to support material delivery need at different stages of projects.

1:10 pm Close of Cell Line Engineering and Development