Cambridge Healthtech Institute’s 8th Annual
Higher-Throughput Protein Production and Characterization
Improving Processes
January 17-18, 2019
High-throughput processing has come of age by transforming the traditional protein-by-protein trial-and-error approach for testing criteria and scaling up. In CHI’s Higher-Throughput Protein Production and Characterization conference, HTP is explored
in the quest to develop methods that ensure quality and translate to large scale much more quickly and efficiently than in the past. Automation, robotics and liquid handlers will be discussed, along with developing small-scale models that shed light
on bioproduction. Case studies will be presented that illustrate how leaders in the field are integrating HTP approaches to reduce the time and effort needed to successfully analyze proteins, fine tune processes, and achieve well-folded, pure protein.
Final Agenda
THURSDAY, JANUARY 17
7:45 am Registration and Morning Coffee (Sapphire West Foyer)
8:10 Organizer’s Welcome Remarks
Mary Ruberry, Senior Conference Director, Cambridge Healthtech Institute
8:15 Chairperson’s Opening Remarks
Kelcy Newell, PhD, Senior Scientist, Laboratory Automation & High-Throughput Process Development, MedImmune, LLC
KEYNOTE PRESENTATION
8:20 Downstream Processing in Biomanufacturing: Multimodal Chromatography, Affinity Precipitation and Integrated Bioprocessing
Steven Cramer, PhD, William Weightman Walker Professor, Isermann Department of Chemical and Biological Engineering, Rensselaer Polytechnic Institute
Targeted experiments and molecular simulations will be used to shed light on the importance of protein surface clusters and ligand properties for creating selective separations in multimodal chromatography. Affinity precipitation using smart biopolymers
for the simultaneous recovery and purification of both mAb and non-mAb biologics will then be presented. Finally, results will be given on a novel approach for the rapid development of integrated downstream biomanufacturing processes for biological
products.
9:00 Platformization of Multi-Specific Protein Engineering II: From Automated Transfection to High-Throughput Multi-Parametric Characterization of Large Variant Libraries
Joerg Birkenfeld, PhD, Section Head, High Throughput Biologics, R&D Biologics Research/Protein Therapeutics, Sanofi-Aventis Deutschland GmbH
The success rate to identify a multi-specific lead molecule with favorable drug-like properties increases with the number of engineered variants tested. We recently established a novel, fully automated platform process for the in silico design and fast generation of large panels of multi-specific variants. Here, we report on the integration of miniaturized lab unit operations with cutting-edge automation for transient transfection, expression, purification and characterization
of up to 10,000 engineered variants in high-throughput.
9:30 POSTER SPOTLIGHT
Opus RoboColumn Lifetime Studies, Failure Modes, and Lifetime Extension Methods to Enable Biopharmaceutical Manufacturing Processes
Briana Baldi, MSc., BioProcess Engineer, Downstream Technology Transfer, MedImmune, LLC
10:00 Coffee Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
11:00 Applications of Modular Expression Toolboxes in High-Throughput Protein Expression
Ernst Weber, PhD, Laboratory Head, Biologics Lead Optimization,
Project Leader, Ophthalmology, Bayer HealthCare
The presentation will focus on the setup of a modular expression toolbox, consisting of standardized elements influencing expression levels, which allow the rapid generation of multiple expression constructs and also the generation of complex
expression optimization libraries. Advantages and implications of a modular cloning system including implementation into protein expression optimization workflows will be discussed and a number of successful case studies will be presented.
11:30 Self-Cleaving Tags Based on Split Inteins: Increased Reliability Enabling Higher-Throughput Applications
David Wood, PhD, Professor, Chemical & Biomolecular Engineering,
The Ohio State University
An important limitation of intein-based self-cleaving tag systems is a lack of reliability for arbitrary target proteins. In some cases, the intein tags cleave too quickly, while in others the tags cleave too slowly or not at all. In our recent
work, we have developed several systems to interrogate the sources of rate variations, and can now provide detailed guidance on design and operation of these methods in higher-throughput applications.
12:00 pm Session Break
12:10 Luncheon Presentation (Sponsorship Opportunity Available) or Enjoy Lunch on Your Own
1:10 Ice Cream Break in the Exhibit Hall with Poster Viewing (Sapphire Ballroom)
2:15 Chairperson’s Remarks
Edward Kraft, PhD, Senior Scientific Manager, BioMolecular Resources, Genentech, Inc.
2:20 High-Throughput Mid-Scale Antibody Purification and Characterization
Jiansheng Wu, PhD, Senior Scientist, Protein Chemistry, Genentech, Inc.
Recent advances of high-density mammalian cell culture dramatically increases the titer of transiently expressed antibody. Several new transient systems, such as Expi 293, express antibody at 300-500mg/L, makes it possible to use 100-200ml
cell culture to produce 20-50mg of antibody, which is sufficient for many in vitro and in vivo studies. To fully take advantage of these systems, we have developed a high-throughput purification and characterization platform
for mid-scale antibody production, which can perform automated two-step purification from 30-500ml culture. The integrated system enables high-throughput liquid handling, automated analytical SEC and mass spec analysis, and batch uploading
of QC samples into database.
2:50 Nanodelivery of a Functional Membrane Receptor to Manipulate Cellular Phenotype
Matthew Coleman, PhD, Senior Scientist, Physical
Life Sciences, Lawrence Livermore National Laboratory (LLNL)
We have developed a platform that enables multiplex investigation of G-protein-coupled receptors (GPCRs) by coupling cell-free expressed GPCR in E. coli with functional profiling at the single-molecule level for
delivery of the active GPCR into mammalian cells. Specifically, our work shows that we can assemble full-length, wild-type β2AR associated with nanolipoprotein particles (NLPs) in cell-free E. coli lysates
in a single step process. We then functionally characterized the nanoassemblies by demonstrating ligand-induced confirmation activation.
3:20 POSTER SPOTLIGHT
Semi-Automated Medium Scale High Throughput Magnetic Bead Purification
Vivian Li, Scientist, Molecular Structure, Amgen, Inc.
3:35 Networking Refreshment Break (Sapphire & Aqua West Foyer)
4:00 Bridging the Silos: Integration of HTPD Scale-Down Models and High Throughput Analytics Enables and Accelerates Process Development of Biopharmaceuticals
Kelcy Newell, PhD, Senior Scientist, Laboratory Automation
& High-Throughput Process Development, MedImmune, LLC
As biopharmaceutical companies shift their pipeline to increasingly novel therapeutics, development is filled with new challenges including different novel product quality attributes, new product and process related impurities, and/or accelerated
product degradation pathways. We have adopted a cross-functional approach to minimize disruption of development timelines and even enable acceleration to market when required. This talk will spotlight multiple cross-functional high-throughput
process development strategies that have been successfully utilized for problematic biopharma molecules in development.
4:30 A Computational High-Throughput Method for the Study of Modified RNA Interactions with Proteins
Phanourios Tamamis, PhD, Assistant
Professor, Chemical Engineering, Texas A&M University
Little is known about the abundance of protein-RNA modification interactions and how these interactions may regulate protein function. Here, we present the first, to our knowledge, computational protocol for the characterization of interactions
between proteins and RNAs containing post-transcriptional modifications. As an initial test case, we implemented our CHARMM-based protocol to investigate interactions between E. coli polynucleotide phosphorylase
protein with modified RNAs, demonstrating a reasonably high agreement between computational and experimental results.
5:00 CANCELLED: Application of Native MS for the Characterization of Bispecific Antibodies during Drug Development
Markus Haberger, PhD, Senior Scientist, Pharma
Technical Development Analytics Extended Characterization, Roche Diagnostics GmbH
High-molecular weight aggregates, such as antibody dimers and other side products derived from incorrect light or heavy chain association, typically represent critical product-related impurities for bispecific antibody formats. In this study,
an approach employing ultra-pressure liquid chromatography size-exclusion separation combined with native electrospray ionization mass spectrometry for the simultaneous formation, identification and quantification of size variants in recombinant
antibodies was developed. Samples exposed to storage and elevated temperature(s) enabled the identification of various bispecific antibody size variants.
5:00 Close of Day
FRIDAY, JANUARY 18
8:00 am Registration (Sapphire West Foyer)
8:00 BuzZ Sessions with Continental Breakfast (Sapphire Foyer)
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.
Click here for more details
Moderator: David Wood, PhD, Professor, Chemical & Biomolecular Engineering, The Ohio State University
CANCELLED: FEATURED PRESENTATION
9:05 Medium Scale (0.25-10L) High-Throughput Paramagnetic Purification of Biologics
John Kawooya, PhD, Director, Biologics Optimization, Discovery Research, Amgen, Inc.
Here, we describe a transformative medium-scale rare earth magnetic (NdFeB) system which purifies biologics directly from crude cell culture with cells. The capture step on the beads starts 18-24 hours before the end of protein expression,
thereby eliminating the cycle time traditionally spent during centrifugation, clarification and sample loading. The output of the system is amplified by formatting and magnetizing sixteen tanks each capable of purifying more than 2
grams of protein in less than two hours.
9:30 Chairperson’s Remarks
Phanourios Tamamis, PhD, Assistant Professor, Chemical Engineering, Texas A&M University
9:35 High-Throughput Purification of Synthetic Peptides
Mathias Schaffrath, PhD, Group Head, R&D IDD In vitro Biology & HT Chemistry Library, Chiral & Peptide Purification, Sanofi-Aventis Deutschland GmbH
The purification of synthetic peptides is still a challenge. Reversed phase chromatography is in many cases the method of choice. Sometimes orthogonal reversed phase methods with two chromatographic steps and two different column selectivities
are needed to increase the purity to more than 95%. Chromatographic experience, a thorough method development and up scaling is needed for successful separations. Partial automation of the process leads to remarkable throughput, which
is particularly important in the field of research.
10:05 Evolving the High-Throughput Protein Purification Pipeline
Edward Kraft, PhD, Senior Scientific Manager, BioMolecular
Resources, Genentech, Inc.
The development of high-throughput protein expression and purification pipelines are an essential component for predicting construct design success and scalability for protein production. This process requires significant expertise spread
across biochemistry, biology, automation and informatics to create a system that has the flexibility to impact all types of proteins. This talk will present our work to continually adapt our high throughput protein production workflows
to support the diversity of non-antibody proteins in support of research across Genentech.
10:35 Networking Coffee Break (Sapphire & Aqua West Foyer)
11:00 SPEAKER CHANGE: Library-Based Glycan Identification by Mass Spectrometry in Combination with Fluorescence Quantification as a Biopharma Solution for Automated Glycan Characterization
Robert Wilmanowski, PhD, Director, Instrumental Bioanalytics, Glycotope GmbH
In the present study, proteins comprising different numbers of glycosylation sites were analyzed by release of N-glycans with N-glycanase F, fluorescence labeling of N-glycans, data recording by use of HILIC-UPLC-FLD-ESI-QTOF MS/MS (hydrophilic
interaction ultra-performance chromatography with fluorescence detection coupled to electrospray ionization quadrupole time-of-flight tandem mass spectrometry). Subsequently, automatic data processing was performed, and final reporting
of all data in a certificate of analysis.
11:30 Beyond Miniaturization and Parallelization: Standard and Tailor-Made Automated Workflows for Smart Microbial Phenotyping and Bioprocessing
Marco Oldiges, PhD, Professor and Head, Bioprocesses
and Bioanalytics, Institute of Bio- and Geosciences, IBG-1, Biotechnology, Forschungszentrum Jülich GmbH
Microbial production of heterologous proteins demands increased cultivation throughput at well-defined bioprocess conditions. Making use of miniaturization, parallelization and automation, standard and tailor-made workflows need to be
put in place, comprising the full experimental pipeline from upstream processing, cultivation, process analytics, data management and design-of-experiment. Case studies illustrate how developments in miniaturized cultivation combined
with smart lab automation and data processing are amalgamated in workflows for more efficient microbial phenotyping and bioprocess development.
12:00 pm Conference Wrap-Up
David Wood, PhD, Professor, Chemical & Biomolecular
Engineering, The Ohio State University
12:30 Close of Conference