We will describe and evaluate different process intensification strategies for the mAb capture step. Strategies will depend on mode of upstream operations (fed batch or perfusion) and the desired process outcome of the capture step, i.e. improved productivity, and cost or buffer consumption. We will discuss opportunities with the recent introduction of high capacity protein A resins, how and when multicolumn chromatography is beneficial and how emerging technologies such as rapid cycling Fibro chromatography can address bottlenecks.

Topic areas that could align:

• Development and Implementation of Next Generation Downstream Processes
• Novel Purification Technologies

Speaker:

Gunnar Malmquist, Ph.D
Principal Scientist
Cytiva

Clinical and commercial continuous manufacturing of biologicial products has taken another step forward with the successful production of monoclonal antibody from our fully disposable 500 L manufacturing scale non-GMP MaruX^TM facility. Using the ApolloX^TM high biomass perfusion cell-line with commercially available upstream systems and SymphonX^TM purification skids for point-of-use buffer dilution and connected, intensified batch processing we were able to process ~1 kg mAb/day into downstream.

Speaker:

Charles Heise
Senior Staff Scientist, Bioprocessing Strategy & Development group
FUJIFILM Diosynth Biotechnologies

A brief introduction into mechanistic modeling of preparative chromatography, how fundamental natural laws can be employed to interpret complex data, understand the physicochemical phenomena and simulate processes for purposes of optimization and characterization. This talk will focus on how mechanistic modeling is especially beneficial for novel formats and their downstream processes, in which data may be sparse, variable or non-existent, material supply may be lacking and process or analytical variability is often problematic. By using experimental data with unparalleled efficiency, a mechanistic model enables one to interrogate a process in silico, enabling complete flexibility in evaluating process conditions, extrapolating outside of calibration data and ultimately generate process understanding.

Speaker:

Dr. Thiemo Huuk
Chief Executive Officer
GoSilico GmbH

• How Biogen currently streamlines downstream process development activities with high throughput and automation tools. An overview of the latest improvements we have added and how to increase efficiency
• What we are looking into to further help expedite workflows: High throughput TFF system, multi-column small-scale processing
• A case study demonstrating efficiency gains

Speaker:

Juan Cueva Tello
Scientist, Process Biochemistry
Biogen

In the face of increasing global demand for insulin product, there is a pressing need to develop a more efficient and economical process for the production of recombinant insulin therapeutics. Here, we present a new and efficient downstream processing workflow for recombinant human insulin production using E. coli as the host system. For sample preparation prior to chromatography purification, Bio-Gel P-4 was identified as a very efficient resin for proinsulin desalting; for proinsulin purification, capture with Nuvia S AEX resin and elution in sodium citrate buffer achieved both good recovery (91-96%) and purity (81–84%); and for insulin purification, Nuvia HR-S AEX resin was found to be highly effective in removing a significant amount (~90%) of higher molecular weight cleavage product impurities. This new chromatographic process may serve as an excellent chromatographic tool for the purification of recombinant human insulin produced from E. coli Inclusion Bodies.

Overview

• Insulin and proinsulin purification workflow
• Effects of pH and conductivity
• Resin characteristics and optimization

Speaker:

Zhang Wei, PhD
Staff Scientist and DSP Group Head, Downstream Processing Group
Bioprocessing Technology Institute of Singapore at A*STAR

Mark A. Snyder, PhD,
Manager, Process Chromatography, R&D Applications Group
Bio-Rad Laboratories

Downstream processing involves multiple unit operations including filtration and several chromatography steps that incrementally increases the purity of the target by exploiting the physical and chemical properties. It is critical for therapeutic monoclonal antibodies to have very high purity with low process related impurities like host-cell proteins (HCPs), DNA, leached protein A and aggregates in order to be administered to patients safely. We have identified a novel approach to improve impurity clearance efficiency of chromatographic steps by modulating the interaction of target protein and/or impurities with ligands on the resins. Protein A resin with high capacity (PROchievATM) or mixed-mode HIC resin (Bakerbond^R Poly Hi-propyl) is used for a capture step and additives are screened to reduce the non-specific binding of impurities. Following IEX steps are optimized to increase the capacity and removal of impurities such as residual HCPs, DNA, charge variants, aggregates and leached protein A.

Moderator:

Speaker:

Jungmin Oh
Manager - New Product Development
Avantor

Recently the requirements of virus filtration are changing. For example, the protein concentrations of process solutions are increasing, and the virus filtration step is operated not only with pressurized vessels but more and more with pump-based system. In this 2-part presentation, we discuss about “Advanced Virus Filtration – High Protein Concentrations and Pump-Based Virus Filtration”.

Increasing cell culture productivity and improving column chromatography performance in the past several years have resulted in virus filtration feeds with higher protein concentrations. Feed solutions at higher protein concentrations can present some challenges for example the potential increase in aggregation formation which in turn can affect filterability. However, one main benefit at higher concentration is the reduced filtration surface area needed and therefore the cost, providing that the nanofilter can handle this increase in concentration. Other benefits include reduced processing time and reduced footprint. Planova BioEX exhibits high throughput with high concentration mAbs and the addition of Virosart Max prefilter can further improve filterability by removing aggregates.

Viral clearance validation studies are typically performed using small-scale filters operated by applying constant pressure to a feed vessel, while at large-scale it is favored to use pump-based systems utilizing single-use bags, which offer improved footprint and portability. Pump-based filtration processes present more process complexity, more potential for process fluctuations, and more challenges in scaling. We have performed studies to better understand pump-based virus filtration operations using Planova™ filters. We show that Planova 20N filters provide effective viral clearance even in the presence of large fluctuations in transmembrane pressure across all filter sizes and obtain comparable results with that of using constant pressure.

Speaker:

Konstantin Agolli
Senior Product Manager & BioOptimal TFF Specialist
Asahi Kasei Bioprocess Europe

• Lack of coordination in the Upstream to Downstream handoff in process development can result in reduced efficiency and longer development timelines. The use of certain tools during the handoff can accelerate the start of development
• Rapid titer technologies already in use by Upstream enables early product mass prediction and real-time troubleshooting in downstream process development.
• Harvest DBC using variable pathlength technology enables a rapid start to development and generates accurate and reproducible DBC results without analytical support.

Speaker:

Thomas Lindsey
Senior Associate, Downstream Process Development
KBI Biopharma

The BioSMB platform based on the principle of multi-column chromatography provides an ideal opportunity to start exploring possible starting points. For customers seeking lower production costs with a single-use solution for clinical and commercial processing, BioSMB can reduce resin usage by up to 80% with minimal disruption to the accepted ways of doing chromatography today. This presents a considerable saving on operational expenditure, particularly when considering affinity capture steps.

In this virtual event, our Technology Expert Jason Forte, who has over five years of experience in helping customers set up the BioSMB platform in a field application specialist role and enabling them to design their intensified processes, will demonstrate the ease of installation of the single-use flow path and how it facilitates scale-up and walk through the automation software.

During this session you will learn:

• A demonstration of the ease of installation, set-up, and main features of the BioSMB PD and Process systems.
• How the software enables user-friendly processing.
• Valve technology, flow path configuration, and consistent recipe handling between benchtop and GMP scale systems ensuring scalability.
• How the BioSMB system offers a 3-5 fold increase in productivity
• How to yield up to 80% savings on chromatography resin costs with minimal disruption to the accepted ways of doing chromatography today

Speaker:

Jason Forte
Technology Expert, Chromatography Systems
Sartorius

The increasing diversity and complexity of new advanced therapy medicinal products (ATMPs), with unique and non-standard technology approaches, is creating challenges in the biomanufacturing industry. The lack of downstream processing platforms that can accommodate this diversity often results in long processing times with high loss of yield. Custom affinity ligands are designed for specific applications and can therefore significantly improve bioprocessing economics by reducing the number of process steps and increasing the capture and recovery of ATMPs.

This presentation outlines strategies for ligand design and screening. We discuss the ligand discovery process, considering approaches for the information available, such as the target protein structure, binding site and known ligands or inhibitors. Additional factors such as stability, yield and cost will also determine the choice of ligand library, so the presentation includes the following topics:

• Understanding the ligand discovery process
• The advantages of using synthetic ligand libraries
• How Affimer® ligand technology offers unique solutions for affinity purification

Speaker:

Eric Bugglin-Borer
Separations Product Specialist,
Astrea Bioseparations

• Manufacturing processes for antibodies have evolved significantly over last several decades
• While platforms are reasonably well established, there are several new challenges still exist
• This talk will highlight the latest key problem areas in our industry for downstream development and strategies for handling them
  • High mass coming from higher titers
  • mAb reduction during harvest
  • PS80 degradation in DS
  • High concentration DS
  • Improving throughput
  • Newer and more complex mAb formats

Speaker:

Sanchayita Ghose, Ph.D.
Executive Director, Head, Global Downstream Bioprocess Development
Biologics Development, BMS

Chimeric constructs constituting complex products often have compromised solubility and stability characteristics. An automated workflow has been developed to rapidly identify process conditions and product concentrations complex products can be manufactured at. As a result protein solubility data can be generated in a broad range of buffer and excipient compositions using low amounts of material. The data can be leveraged to inform chromatographic purification study design

Speaker:

Rhesa Budhidarmo
Senior Scientist R&D
Lonza

Host cell proteins (HCPs) are one of the major part of process-related impurities during manufacturing of protein-based biopharmaceuticals. They are considered a critical quality attribute (CQA) due to their potential immunogenicity and impact on product stability. Typically HCPs are routinely monitored by ELISA commercial kits, whose performance primarily relies on the capability of the employed anti-HCP antibodies to cover HCPs. This method is reliable for early phases projects but does not provide a full coverage of the total population of HCP. In addition, ELISA method can be time consuming and laborious. Since in Lonza IBEX we use Xceed cell line as a platform for Process development, we generated antibodies against HCPs of this specific cell line to reach an optimal and precise quantification of HCP impurities. We developed an HCP quantification method using the Gyrolab® xPand as a semi-automated platform technique. With specific Gyrolab Bioaffy™ CDs and method generated by Gyros Protein Technologies, we obtained high working range and high sensitivity in the low-concentrated part of the standard curve. Finally, we compared performance results in HCP quantification between ELISA and Gyrolab xPand methods showing similarity for most of our Downstream in-process samples.

Speaker:

Sottas Valentin
Head of Purity and Impurity Team, Analytical Development
Lonza

Shorter timelines, larger data requirements. How can you balance the need to reduce time to experiment, time to clinic, and time to market while generating data comprehensive enough to avoid extended review?

Mechanistic models offer a way to negotiate a rapidly diversifying pipeline of molecules. Mechanistic models — mathematical interpretations of physiochemical interactions in chromatography — are already making a profound impact on the workflow of process development.

In this talk, we will review in-silico process development and data-driven decisions to see how mechanistic models can help you reduce timelines and improve outcomes with greater predictive power, both within and outside of the explored characterization space.

Speaker:

John Scibetta
Advanced Chromatography Specialist
Cytiva 

The risk for transmission of emerging viruses is serious threats to public health and have become a global concern. As a leading company that launched the world's first virus removal filter Planova, Asahi Kasei has contributed to the virus safety of pharmaceutical products through collaboration with various pharmaceutical companies, academic institutions and regulatory agencies.

This presentation introduces the history and progress of our challenge to emerging viruses.

Speaker:

Sebastian B. Teitz, Ph.D.
Scientific Coordinator
Asahi Kasei Bioprocess Europe, Cologne Technical Center