Cambridge Healthtech Institute’s Second Annual

Bioprocess Analytics

Data Measurement, Monitoring and Modeling Allow Informed Control of Bioprocesses

January 11-12, 2018

 

The biopharmaceutical industry is meeting increasing demands and costs for biotherapeutics through process optimization. Advanced instrumentation with sampling techniques, new sensor technologies and analyzers have emerged to monitor both upstream and downstream processes. These analytical tools, however, result in large, complex datasets with multivariate interactions. The inherently complex nature of these datasets makes extraction of meaningful and relevant information a difficult task.

Cambridge Healthtech Institute’s Second Annual Bioprocess Analytics conference addresses statistical analysis strategies including multivariate data analysis (MVDA), quality by design (QbD), process analytical technology (PAT) and multi-attribute method (MAM), allowing for optimized and informed control of bioprocessing.

Final Agenda

THURSDAY, JANUARY 11

7:45 am Registration and Morning Coffee

Process to Product

8:15 Chairperson’s Opening Remarks

Gyun Min Lee, Ph.D., Professor, Biological Sciences, KAIST

KEYNOTE PRESENTATION

8:20 Integrating Cell Culture with Magnetic Protein A Bead-Trap Technology Accelerates Antibody Purification

John_KawooyaJohn K. Kawooya, Ph.D., Director, Biologics Optimization, Discovery Research, Amgen

Antibody engineering produces large numbers of molecules (200-500 molecules at 30-50ml each) which require purification, analysis and screening for potency, binding, pharmacodynamics, pharmacokinetics, manufacturability, expression levels and stability in order to select leads. Ever since its inception over 30 years ago, the AKTA system combined with Protein A agarose columns has remained the “workhorse” of antibody purification from cell cultures. However, the inability of this system to process multiple samples in parallel coupled with both its limiting flow rates, its requirement for multiple FTEs to remove cells and particulate from each sample prior to loading together with the potential for sample swapping errors and cross contaminations – all impose major bottlenecks in expediting large purified panels of molecules. In this presentation, I show how a single FTE with parallel Magnetic Protein A bead-trap technology accelerates delivery of high-quality purified antibodies in high yield directly from small (30ml-5liters) to large (25-liter wave-bag) crude cell cultures without centrifugation or filtration.

9:00 Speed to IND: Alignment and Acceleration of Critical Early Phase Activities

Kyle_ZingaroKyle Zingaro, Ph.D., Development Scientist II, Early Stage Development, Alexion Pharmaceuticals

Speed to IND is the current battle cry across early phase biologics development. Although this faster-is-better approach comes with some risk, new technologies and alignment of workflows can afford both faster and better decisions during this crucial phase of new product development. This is especially true across the Discovery and Process Development handoff including the interface of discovery, preclinical, process, formulation, and analytical teams. Here we present new data and approaches to improve that handoff and detail the impact on timelines and quality of molecules and cell lines in early phase development.

9:30 "Lost in Translation": Bridging the Gap between Academia and Biotech

Tsafi_DanieliTsafi Danieli, Ph.D., Director, BioGiv Excubator & Head, Protein Expression Facility, Wolfson Centre for Applied Structural Biology, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem

One of the most difficult and frustrating aspects of basing a startup company on academic findings is translating and transferring academic findings to biotech language. This procedure is often frustrating to both parties and requires psychological skills as well as critical review of the research. Working in the interphase between academia and industry, there are several preemptive strikes we can take to avoid some of the major pitfalls.

10:00 Coffee Break in the Exhibit Hall with Poster Viewing

11:00 Proteomic Analysis of Host Cell Protein Dynamics in the Culture Supernatants of Therapeutic Protein-Producing CHO Cells

Gyun_Min_LeeGyun Min Lee, Ph.D., Professor, Biological Sciences, KAIST

Host cell proteins (HCPs) accumulate extracellularly during the cultures of recombinant CHO (rCHO) cells, potentially impairing product quality. HCPs accumulated extracellularly in batch and fed-batch cultures of rCHO cell lines were identified and quantified by mass spectrometry. This dataset of HCPs provides insights into determining the appropriate target proteins to be removed during both the cultures and purification steps for ensuring good therapeutic protein quality.

11:30 Using SUREscan™ to Survey Genetic Changes in Stable CHO Cell Lines

Pierre-Alain_GirodPierre-Alain Girod, CSO, Selexis

CHO cells are the most frequently applied host-cell system for industrial protein therapeutic manufacturing. Rapid generation of high-producing clones that don’t lose expression capability over time has been a major industry focus. Using SUREscan™ with next-generation sequencing (NGS), we can quickly analyze whole genomes of any cell line, improving traceability of Research Cell Banks (RCBs). In contrast to other CHO published data, we will show that SUREtechnology Platform™ generates RCBs with chromosomally stable lineages.

12:00 pm Session Break

 Unchained Labs 12:15 Luncheon Presentation: Get Your High Protein Concentrations Right on the Money with Lunatic

Thomas Martens, Principal Scientist, Unchained Labs

Stop quantifying proteins one by one hoping that your old reader is getting the numbers right. Get rid of that dilution step you always need to measure 200 mg/ml IgG or even higher. Come learn about how Lunatic gets rid of all dilutions, eliminates any risk of cross contamination and accurately measures protein concentration at high throughput and high concentrations. We’ll talk about how lunatic: • measures either 16 or 96 samples in one run • uses only 2 µL for each measurement • needs only 5 minutes • Requires no dilutions

1:15 Ice Cream Break in the Exhibit Hall with Poster Viewing

Analytical Workflows and Modeling

2:00 Chairperson’s Remarks

Tsafi Danieli, Ph.D., Director, BioGiv Excubator & Head, Protein Expression Facility, Wolfson Centre for Applied Structural Biology, Alexander Silberman Institute of Life Sciences, The Hebrew University of Jerusalem

2:05 Novel Hybrid Modeling Approaches for QbD: Getting More from the Combination of Fundamental Knowledge and Statistical Methods

Moritz_von_StoschMoritz von Stosch, Ph.D., Senior Manager, Fermentation, Technical R&D, GSK Vaccines

The combination of fundamental knowledge with statistical methods is referred to as hybrid modeling. These models have been shown to 1) provide better extrapolation properties, 2) have lower data requirements, and 3) are more efficient to develop than models based on a single knowledge source. This talk introduces novel hybrid methods that work more coherently with small datasets and estimate their own prediction quality, such being optimal for the development of the process design space in the context of QbD.

2:35 Phase-Appropriate Analytics

SiowFong_WeeSiowFong Wee, Ph.D., Director, Formulation, Analytical & Bioassay, Aptevo Therapeutics

It can be a struggle to decide how much analytics is considered sufficient for product quality evaluation and characterization to support a project that is in the early development stage. Phase-appropriate analytics to support ‘clone-to-clinic’ will be presented in this talk.

3:05 Integration of ambr High-Throughput Bioreactor Systems into the USP Development Workflow and into the Data Acquisition, Management and Analysis System

Timo_FrensingTimo Frensing, Ph.D., Senior Scientist, Cell Culture Research, Roche Diagnostics GmbH

To enable a high-throughput USP development we implemented a semi-automated workflow to connect ambr® bioreactors (Sartorius, Germany) via a Fluent® pipetting robot (Tecan, Switzerland) to the Cedex Bio HT Analyzer (Roche Diagnostics, Germany) for an accurate sample processing and sophisticated process analytics. Thereby, all systems are integrated in our data acquisition, management and analysis system to ensure an efficient data processing.

3:35 Refreshment Break in the Exhibit Hall with Poster Viewing

Cell Line Development

4:15 Lost in Translation: On the Formation of Protein Sequence Variants

Zhongqi_ZhangZhongqi Zhang, Ph.D., Scientific Director, Attribute Sciences, Process Development, Amgen

With modern mass spectrometry and appropriate informatics tools, a large number of low-level sequence variants in therapeutic proteins are detected and quantified. This large collection of information allows for a deeper understanding of the mechanism for the formation of sequence variants, thereby facilitating optimization of cell line and cell culture process to minimize them.

4:45 The Stability of CHO Genome: Essential for Cell Line Characterization or Not?

Noriko_YamanoNoriko Yamano, Ph.D., Senior Scientist, Manufacturing Technology Association of Biologics; Guest Academic Staff, Graduate School of Engineering, Osaka University

The chromosomes in CHO cells frequently cause genomic variations, due to genetic instability. Distribution and stability of chromosomes were examined in CHO-DG44 cells, and two cell lines expressing different numbers of chromosomes were isolated from the original CHO-DG44 cell line to investigate the effect of aneuploid cells on recombinant protein production. In addition, gene expression profiles between cells with disparate chromosome numbers have been compared by mRNA-seq analysis.
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5:15 High-Throughput Screening of Transfection Efficiency of dTtaPS Reagent Library, and Its Application for Transient Production of Proteins in Micro Bioreactors

Harsh Jain, Ph.D., Visiting Associate, FDA

5:45 Close of Day

FRIDAY, JANUARY 12

8:00 am Registration

8:00 BuzZ Sessions with Continental Breakfast

Protein therapeutics is a fast-growing global market. As the science improves, so does the complexity of the R&D organization. Ensuring product quality plus speed to market requires insights from stakeholders working across the stages of protein science R&D. Join experts representing this PepTalk pipeline, peers, and colleagues for an interactive roundtable discussion. Topics include highlights from the week’s presentations, new technologies and strategies, challenges, and future trends.

Table Moderator: Diane Paskiet, MS, Director of Scientific Affairs, Scientific Affairs and Technical Services, West Pharmaceutical Services

Table Moderator: Moritz von Stosch, Ph.D., Senior Manager, Fermentation, Technical R&D, GSK Vaccines

 

Product Characterization and Analytics

9:00 Chairperson’s Remarks

Moritz von Stosch, Ph.D., Senior Manager, Fermentation, Technical R&D, GSK Vaccines

9:05 Characterization of mAbs Using Charge Variant Analysis Coupled to High Resolution Native Mass Spectrometry

Jonathan_BonesJonathan Bones, Ph.D., Principal Investigator, Characterisation and Comparability Laboratory, National Institute for Bioprocessing Research and Training

Charge variant analysis (CVA) of biopharmaceuticals is required under ICH Q6B. Issues arise when a new peak is identified in the CVA profile. Here, the development of high resolution charge variant analysis coupled directly to native high resolution mass spectrometry is described that facilitates the intact mass analysis of minor charge variants with high mass accuracy. Application to the characterization of mAbs and other recombinant proteins is described under normal conditions and during forced degradation studies.

9:35 Utilizing High-Throughput Lab Automation and Analytics, Advanced Data Management and Multivariate Statistical Analysis to Define the Formulation Design Space for Biotherapeutics

Michael_SiedlerMichael Siedler, Ph.D., Head, NBE High-Throughput and Advanced Formulation Sciences, Drug Product Development, AbbVie Deutschland GmbH & Co. KG

We discuss adaption and miniaturization of standard analytical methods to be used for 96 and 384 well plates, transition to a data-centric strategy, and implementation of advanced data management to effectively integrate and analyze screening data (including metadata). We also discuss multivariate parameter analysis and statistical modeling to calculate the formulation design space, hence assuring safety and efficacy of new products.

10:05 The Use of Method Performance Monitoring as a Component of a Comprehensive Product Control Strategy

Juma_BridgewaterJuma Bridgewater, Ph.D., Senior Scientist, Analytical Sciences and Technology, Bristol-Myers Squibb

A pharmaceutical product control strategy is designed to ensure that final product quality is representative of product shown to be safe and efficacious. This is not just controlled by testing to specifications but is the result of a comprehensive system of process controls such as raw materials, intermediates and process parameters and analytical testing. High-quality assays must be used for each of these controls and there is a thorough assessment of method performance and robustness when methods are implemented and control limits are defined. However just as for process performance, method performance can vary over time. The use of real-time method performance monitoring supports a comprehensive product control system by providing an assessment of the quality of the data being generated to assess process performance and product quality. This additional dimension to product control can be critical for the early identification of issues that may mask assay failures or yield false positive results that may otherwise go undetected. The collection of this data over time also supports more effective assay and process troubleshooting and the redefinition of assay controls to improve assay robustness.

10:35 Coffee Break with a Poster Pavilion

PepTalk is proud to support and recognize the protein scientists of tomorrow during the Poster Pavilion. This time has been set aside to view the Student Fellowship posters and interact with presenters one on one. This opportunity gives job seekers the chance to share their expertise with future/potential employers or develop contacts to further their research.

Higher-Throughput Systems

11:15 High-Throughput Automations and Optimizations for Improved Binder Generation and Validation

Jonas Schaefer, Ph.D., Head, High-Throughput Binder Selection Facility, Biochemistry, University of Zurich

While recombinant binder selection pipelines by now work in rather high-throughput, the screening of suitable affinity reagents and especially the validation of their essential features for the final applications is still laborious and time-intensive. To optimize the efficiency of these processes, we have improved already existing and developed novel methods to efficiently test candidates for their suitability, e.g., regarding their specificity.

11:45 High-Throughput Characterization of Hydrolytic Enzymes in Low Volume and Closed Systems

Nigel F. Reuel, Ph.D., Assistant Professor, Chemical and Biological Engineering, Iowa State University

Hydrolytic enzymes play a significant role in biologic and synthetic processes. The ability to better characterize these enzymes would enable shorter development times and better products. This talk will detail two recent developments for hydrolytic enzyme characterization: 1) a carbon nanotube-based optical sensor that allows for quantitative measurement in <20ul volumes and 2) a resonant antenna sensor that passively transmits its response in the 1-100MHz range, enabling detection within closed, opaque systems.

12:15 pm Conference Wrap-Up

Richard Altman, MS, Scientist, Protein Technologies, Amgen

Haiyan Jiang, Ph.D., Principal Scientist, Biologics Research, Janssen BioTherapeutics, Janssen R&D

Diane Paskiet, MS, Director of Scientific Affairs, Scientific Affairs and Technical Services, West Pharmaceutical Services

Bjørn Voldborg, Director, CHO Cell Line Development, The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark

Moritz von Stosch, Ph.D., Senior Manager, Fermentation, Technical R&D, GSK Vaccines

 

12:45 Close of Conference