Cambridge Healthtech Institute's 11th Annual

Higher-Throughput Protein Production

Analyzing & Improving Processes

January 17 - 18, 2022 ALL TIMES PST

High-Throughput techniques (HTP) have become essential strategies for improving protein production processes, speeding up results and lowering costs. HTP screening and automation along with the resulting digitalization via these sampling techniques, new sensor technologies, and analyzers are facilitating this productivity. The Higher-Throughput Protein Production conference convenes the innovators who are driving and integrating HTP approaches to reduce the time and effort needed to successfully produce proteins, fine tune processes, and ultimately develop new classes of biological products.

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 400

9:00 am Organizer's Welcome Remarks

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

9:05 am

Chairperson's Opening Remarks

Stefano Menegatti, PhD, Assistant Professor, Chemical & Biomolecular Engineering, North Carolina State University
Sarah M. Rue, PhD, Associate Director, Advanced Automation Technologies, Genomics Institute of the Novartis Research Foundation

Researchers are continuously searching for new ways to improve and accelerate the process of biotherapeutics development. For over 10 years, we have successfully applied automation to accomplish these goals by using our in-house-built Protein Production and Purification Platform (PEPP) to support hybridoma, transient HEK, and transient and stable CHO workflows. Here, we describe PEPP2, a brand new protein production platform. We detail how PEPP2 was designed for increased speed and double throughput compared to the original PEPP system, how new hardware enables automated magnetic bead-based protein purification, and how PEPP2 will be used to build the Secretomics 2.0 protein collection.

9:40 am

3D Printed Convective Media for the Separation of Small and Large Biomolecules

Gregory Dutra, PhD Student, Department of Biotechnology, BOKU

Based on computational fluid dynamics we fabricated precisely ordered three-dimensional chromatography columns with a pore structure relevant for chromatography. The novel material was generated by 3D printing and the advantages for separation of large and small biomolecules, including bionanoparticles like viruses and virus like particles will be presented.

10:10 am

Modeling Approaches in High-Throughput Systems

Stefan Haider, PhD, Scientist, Process Science & Downstream Development, Boehringer Ingelheim RCV GmbH & Co KG

The fully-automated fermentation platform at Boehringer Ingelheim in Vienna has been developed to the point that it can routinely carry out up to 100 fed-batch fermentation runs per week. Huge amount of data is generated, which in turn paves the way for implementing both mechanistic and statistical modeling approaches. Such models help accelerate process development and build knowledge to reduce developmental efforts, ensuring deeper process understanding and better manufacturing processes. In this talk, I will highlight the implemented models used in our high-throughput platform and their uses in different parts of process development. 

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

Accelerating Higher-Throughput Discovery Workflows with Informatics Advances and Next-Gen Predictive Tools

Daniel Yoo, Senior Scientist, Therapeutic Discovery, Amgen Inc.

As biologic therapeutics continue to increase in complexity, innovative approaches to candidate screening, characterization and development are more important than ever. Our approaches to high-throughput protein production incorporate advanced analytics, automation and high-quality informatics to enable robust molecule screening, selection and scale-up. These enhancements enable advances in the speed, quality and productivity of our biologics development pipeline.

11:30 am

In/At-Line Analytics for Rapid Assessment of Titer and Critical Quality Attributes of Therapeutic mAbs

Stefano Menegatti, PhD, Assistant Professor, Chemical & Biomolecular Engineering, North Carolina State University

With dozens of monoclonal antibodies (mAbs) in production, accurate in/at-line mAb quantification is vital to biomanufacturing. Current mAb quantification is time-/labor-intensive. Therefore, we developed the "Dual-Affinity Ratiometric Quenching" (DARQ) assay for rapid quantification of mAb titer and glycosylation, and host cell protein (HCP) titer. The assay is reproducible (variation <1%) and rapid (5 min), and offers excellent sensitivity (<0.5 ng/mL), limit of detection (<100 ng/mL), and dynamic range (100-1600 ng/mL).

Nishant Saxena, PhD, Product Manager, Catalog Product, GenScript

Protein purification using traditional chromatography is limited by throughput and requires labor-intensive sample preparation processes. Magnetic beads-based purification permits the incubation of the beads directly into cell culture or crude lysates regardless of sample volume. This provides a simplified approach to direct target capture while minimizing preparation steps and potentially improving the quality of final product. The tools and their application to simplify protein purification, screening cost-effectively will be described.

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


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 9: Future Platforms for Future Modalities

David W. Wood, PhD, Professor, Chemical & Biomolecular Engineering, The Ohio State University

Scientific breakthroughs in gene therapy and new targets and strategies have stimulated the biopharmaceutical industry to consider highly diverse molecules and delivery methods.  These new approaches will require new manufacturing methods, and like monoclonal antibodies with Protein A, platforms that can quicky bring basic scientific discoveries into the clinic will become critically important.  As we go forward and assemble new information about potential approaches it is worthwhile to stop and examine this progress.  This discussion will be designed to look at what is working and what is not, as well as the pain points and opportunities for the next core platforms that will enable the blockbuster drugs of the future.

  • Are columns here to stay, or is it time to reconsider other approaches?
  • What about cleavable tags?  Is the risk justified by the need for platforms?
  • What kinds of products will be driving this innovation?  What do those platforms need to look like in terms of scale, performance and cost?
  • How open are companies to trying disruptive technologies?  Are the innovations in the products enough for now, so we should play it safe on the methods?​

BuzZ Table 10: BuzZ Session: Automated Cell Culture and Protein Production: Are We There Yet?

Sarah M. Rue, PhD, Associate Director, Advanced Automation Technologies, Genomics Institute of the Novartis Research Foundation
Michelle R. Gaylord, M.S., Senior Scientist II, Genomics Institute of the Novartis Research Foundation

Automated solutions for cell culture and protein production seem to be simultaneously becoming more complex and more critical to drug discovery.  Here, we discuss current challenges with automation, and opportunities for new technology development.

  • How well are your automated cell culture needs being addressed by industry partners?  ​
  • If you could see one new device or platform become available, what would it be?
  • What challenges do you face with automated protein production and/or purification?
  • How could these be addressed?
  • How do you decide whether a process should be automated or continue to be handled manually?
  • How do you handle resistance to implementing automation due to budget concerns, fears about differences in workflow outcomes, etc.?
  • Thinking about available automated incubator technologies, what would you love to see changed or added?


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 400

9:00 am

Chairperson's Remarks

David W. Wood, PhD, Professor, Chemical & Biomolecular Engineering, The Ohio State University
9:05 am

Building Higher-Throughput, Multi-Host, Automated, Mid-Scale, Protein Screening Platform for Drug Discovery

Kanika Bajaj Pahuja, PhD, Scientific Manager, Protein Sciences, Genentech Inc.

We are building higher-throughput, multi-host system, mid-scale, protein expression and screening platform to accelerate drug discovery research at Genentech. This platform is built to perform quick triage of challenging recombinant proteins for structural and biochemical screens. Our semi-automated workflow leverages affinity in tip technology and size exclusion chromatography integrated with liquid robotic handlers to purify multiple samples in parallel. It provides small but sufficient quantities of high-quality proteins to screen for druggability assays and structural determination.

9:35 am

Next-Generation Integrated Antibody Capture Process Based on Magnetic Beads from High Cell Density Cultures

Kristofer Eriksson, PhD, CTO, MAGic Bioprocessing

Cell clarification represents a major challenge for processing suspensions with very high cell densities. By using high-capacity protein A magnetic beads, we have developed an integrated affinity process where cell clarification and antibody recovery is performed in a single step. Using this approach on suspensions with 100 million cells/ml, we achieve adsorptions larger than 95% and yields over 90% with a logarithmic host cell protein clearance of 2–3 and resulting HCP concentration of 5 ppm. Scalability of the magnetic bead-based process from lab to pilot scale is also demonstrated.

10:05 am Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)

Advances in High-Throughput Protein Purification at Different Scales

John K. Kawooya, PhD, Director, Biologics Optimization and Therapeutic Discovery, Amgen, Inc.

Currently, protein engineering is the major gateway to biotherapy discovery.  Protein engineering starts with in silico generation of multi-constructs panels. These constructs are cloned, expressed, purified and analyzed for efficacy and manufacturability. In this workflow, protein purification remains the major bottle neck in accelerating lead identification from multi-construct panels. Here, I present new concepts of the next-generation magnetic and non-magnetic high-throughput parallel systems for de-bottlenecking panel purification.

David W. Wood, PhD, Professor, Chemical & Biomolecular Engineering, The Ohio State University

A practical self-removing affinity tag now exists that can bridge tag-based high-throughput methods for basic research with clinical manufacturing processes to produce tagless products.  Here we describe the development and commercialization of this tag, with a focus on its practicality for a variety of applications at a wide range of production scales.  Case studies demonstrate rapid purification of proteins for research, as well as the potential for large-scale manufacturing of biosimilars and new biopharmaceuticals in the future.

Nigel Shipston, Director - Technical Marketing, FUJIFILM Diosynth Biotechnologies

Lead strain selection is a critical step during microbial process development, initially based on product titer because product quality evaluations are challenged by the timely availability of sufficiently purified material. The capability of a well-proven E. coli expression platform has been enhanced through the use of a scale down/high throughput approach to protein purification using robotics, to enable strain selection based on parallel evaluation of product titer and quality.

12:30 pm Session Break
12:40 pm Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:10 pm Close of Higher-Throughput Protein Production