CMC development webinars
In these live broadcasts, we invite leading developers of new biologics, vaccines, and advanced therapies to share their approaches to CMC strategy, analytical methods, regulatory documentation, and experience with project challenges. The focus of these presentations is on analytical methods for characterizing host cell proteins (HCPs), residual proteins, viral vector proteins, and other proteins present in biologics and advanced therapies.
The invited speakers are all experienced scientists within their respective fields and companies’ products. Upcoming and past events are listed below.
Next event
mAb HCP Control Strategy – Part II:
Experiences from an “off-ELISA” road
Margarita Sabater, Senior CMC Specialist, Genmab, Denmark
17 April 2024 | 16.30 CEST | 10.30am EST | 7.30am PST
In this second Genmab presentation, Margarita will give a detailed overview of how they use LC-MS to assess process changes, demonstrate HCP clearance, and document process consistency.
The presentation includes the following elements for the holistic strategy:
- Process development aided by LC-MS: HCP identification analysis of in-process samples and the bulk mAb product to aid process development and a comparison study of two harvest filter setups to evaluate their efficiency.
- Investigation of problematic HCPs and demonstration of consistent HCP clearance at commercial scale: LC-MS analysis of in-process samples and bulk mAb product from 4 PPQ runs showed consistent removal of HCPs, including problematic HCPs, such as Phospholipase B-like and Carboxypeptidase.
- Feedback from regulatory authorities including EU, Japan, USA, Canada, and UK: Licensing applications in 25 countries with submission data using commercial ELISA kit and coverage analysis by ELISA-MS and LC-MS characterization.
Furthermore, Margarita will discuss the HCP control strategy, including these key issues:
- When should we use mass spec analysis in development?
- What do you do when you detect potentially problematic HCPs?
- What is the future of HCP analytics for mAbs development and perspectives for an MS-only strategy?
Sign up here for the replay of the latest event:
mAb HCP Control Strategy – Part II: Experiences from an “off-ELISA” road
Presenter biography
Margarita Sabater
Senior CMC Specialist
Genmab, Denmark
Margarita Sabater is a Senior Subject Matter Expert (SME) and Analytical Lead in the Late Stage Manufacturing Development group at Genmab. Her group is responsible for developing and validating analytical methods, including potency assays and host cell protein impurity assays, in support of pre-clinical and clinical development of monoclonal antibody programs. Previously, she led the quality control bioassay and impurities group at Novo Nordisk and has wide expertise in host cell impurity analysis. She received her master's (MSc) in chemical engineering from the Autonomous University of Barcelona, Spain.
Past events
mAb HCP Control Strategy - Part I:
Experiences from an “off-gel” road
Søren Skov Jensen, Senior CMC Specialist, Genmab, Denmark
5 March 2024 | Replay available below
The control strategy consists of a list of process inputs and outputs (CQAs) that require monitoring and control to ensure that the process consistently delivers a product of the desired quality. HCPs have traditionally been identified as a critical quality attribute (CQA) that we must control. Among the different ways of controlling HCP levels, release testing by a suitable method and demonstration of process consistency through clearance studies are the most common control elements.
In this presentation, Genmab presents the first elements of their holistic HCP Control Strategy:
- The first step of the HCP control strategy was to obtain knowledge and understanding of the HCP levels in the mAb production process and to identify HCPs in the bulk mAb product using LC-MS. The analysis assessed samples from the Clarified Harvest, Chromatography 1 and Chromatography 2, and the bulk mAb product.
- The second step was to select suitable anti-CHO HCP reagents.
Here, Genmab screened commercial ELISA reagents – 11 kits from 4 suppliers – using the ELISA-MS™ method to investigate which ELISA reagents provided the best coverage of the HCPs present in the process and the final bulk DS.
- In the third step, the commercial ELISA kit that was found optimal for the process and best covered the HCPs present in the final bulk DS was further evaluated by multiple methods for coverage analysis, including a) 2D fluorescence/Western blot, b) Immunoaffinity chromatography (IAC) and 2D DIGE, and c) ELISA-MS.
The webinar concludes with a discussion of future perspectives for HCP analysis, the importance of USP chapter 1132.1, and the change towards MS-based HCP strategies.
Søren Skov Jensen
Senior CMC Specialist
Genmab, Denmark
Søren Skov Jensen is a Senior Subject Matter Expert (SME) in Extended Characterization in the Late Stage Manufacturing Development group, CMC, at Genmab. His group is responsible for extended characterization supporting pre-clinical and clinical development of monoclonal antibody programs. He has a mass spectrometry and proteomics background from the University of Southern Denmark.
Discussion and Q&A with Bryant McLaughlin, Chemistry Manufacturing & Controls (CMC) Executive, USA
7 February 2024 | Replay available below
In this exclusive question-and-answer session, you will hear an industry expert's experiences successfully integrating MS-based HCP analysis into their CMC workflow to optimize process development and ensure regulatory compliance.
You will get the opportunity to gain his input on your concerns and queries to gain clarity on the transformative power of this cutting-edge technology.
Navigate FDA guidelines confidently:
Learn about Bryant's experience with the FDA's - and EMA's stance on Mass Spectrometry (MS)-based HCP analysis. Understand how the technology aligns with regulatory expectations, ensuring your strategies are advanced and compliant.
Learn from real-world experience:
Discover how your peers have utilized the power of MS-based HCP analysis to overcome challenges similar to those you might be facing.
Explore the comparative value of MS-based analysis:
Delve into how MS-based HCP analysis complements traditional methods like ELISAs, including using MS for ELISA HCP antibody coverage determination.
Join and see how MS technology can revolutionize your approach to characterizing host cell proteins and other impurities.
Bryant McLaughlin, PhD
Chemistry, Manufacturing and Controls (CMC) executive, USA
For 15 years, Bryant has held CMC executive positions in biotech companies, including Genentech, Nektar Therapeutics, and 9 Meters Biopharma, developing mAbs and mAb-like products. His extensive experience includes biologics process and analytical development (including residual host cell protein assay validation), process characterization and validation, and global CMC regulatory writing and strategy.
Problematic Host Cell Protein impurities can stop your biologic program!
– Which HCPs to look for and how to measure them
Ejvind Mørtz, Co-Founder & COO, Alphalyse, Denmark
31 October 2023 | Replay available below
Host Cell Proteins (HCP) are process-related impurities created during the manufacturing of vaccines, monoclonal antibodies (mAbs), therapeutic proteins, and viral vector therapies.
Residual HCPs in a drug product can impact patient safety and product stability. In several clinical trials and during the development of many biologics, specific HCPs, such as flagellin, proteases, and the infamous PLBL2, have proved problematic. Some HCPs cause immunogenicity in patients, some HCPS cause unwanted biological effects, and some HCPs can enzymatically degrade the drug itself, decreasing its efficacy and shelf life.
It makes HCPs critical quality attributes (CQAs) that should be measured and documented in all biologics. For mAbs, one of the most well-characterized groups of biologics, it is now well-described which HCPs from the CHO expression system you should be concerned about. Regulatory guidelines assert that the documentation should include risk assessments of individual HCPs identified and removed during purification.
This presentation will introduce you to:
- Examples of bad actor HCPs that have resulted in clinical hold – and even shut down drug development programs
- Descriptions of the main groups of harmful impurities you should watch out for
- LC-MS-based techniques for identification and quantification of HCPs of concern
Ejvind Mørtz, PhD
Co-Founder & COO, Alphalyse, Denmark
Ejvind holds a PhD from the University of Southern Denmark in Protein Chemistry & Molecular Biology. He has more than 20 years of experience developing protein analyses and mass spectrometry methods in the research & development of protein biologics; vaccines, monoclonal antibodies, therapeutic proteins, and cell and gene therapies.
Qualification of a generic CHO HCP assay by orthogonal methods and recommendations for managing critical reagent supply for the validated HCP-ELISA
Robert Hooper, Senior Research Scientist, Rockland Immunochemicals Inc, USA
6 September 2023 | LIVE ONLY
Using a generic off-the-shelf HCP ELISA has several challenges before and after being validated for use in bioprocess monitoring. First, it is difficult to demonstrate that an HCP ELISA is fit-for-purpose to use in bioprocess control strategy, and second consistent long-term reagent supply is an established risk for HCP ELISAs.
To validate HCP reagents, ELISA and 2D coverage are common orthogonal techniques that together demonstrate that the anti-HCP ELISA is fit for purpose. Mass spectrometry provides another way to assess HCP impurities; unlike ELISA, it identifies individual HCP proteins. As such, mass spectrometry is now commonly used as both a primary and an orthogonal method for HCP assessment along with 2D “DIGE/DIBE immunoblots and ELISA.
Here we describe the characterization of a generic CHO HCP assay using standard methods of 2D, ELISA, coupled with an orthogonal LC-MS analysis of the HCP standard and evaluating potential problematic HCPs as part of the qualification. We also propose recommendations for demonstrating fit-for-purpose to the specific bioprocess and for securing long-term HCP reagent supply to reduce the risk of premature critical reagent depletion.
Robert Hooper, PhD
Senior Research Scientist,
Rockland Immunochemicals Inc, USA
Robert has solid experience from both academia and industry, with a strong publication record. Since he joined Rockland in 2019, his primary role has been the development of immunoassays for novel targets, in some instances starting with the validation of newly generated antibodies and taking these through to a finished assay kit for the market.
ELISA reagent characterization using advanced LC-MS methods
Ejvind Mørtz, Co-Founder & COO, Alphalyse, Denmark
28 June 2023 | Replay available below
When using commercial or process-specific ELISAs for Host Cell Protein (HCP) impurity measurements, regulatory reviewers require a characterization of the suitability of the ELISA reagents to the specific product. Our observation from running a vast number of projects, is that frequent challenges in ELISAs are A) Low HCP coverage B) ELISA not measuring abundant HCPs C) lack of dilutional linearity and D) different HCP values obtained when using new ELISA reagents. 2D PAGE analysis cannot answer these challenges, they require higher resolution analytical methods to provide detailed information about the HCP coverage.
In this webinar, you will see results from ELISA reagent characterization using advanced LC-MS techniques on various biologics and expression systems. The analyses include:
- HCP Coverage analysis of early process sample using ELISA immunocapture combined with mass spectrometry (ELISA-MS)
- Coverage of individual HCPs of potential concern
- Coverage of individual HCPs in the purified drug substance
- Comparison of the HCP standard with the early process sample
- Evaluation of high-responding HCPs and non-specific binding likely leading to lack of dilutional linearity
- Evaluation of cross-reactivity to the drug substance
Such a comprehensive ELISA reagent characterization enables a detailed understanding of the HCP-ELISA reagents and documentation of their suitability to the specific manufacturing process and purified drug substance. The LC-MS analyses also help bridging between ELISA reagents and explain differences in HCP measurements between old and new reagents.
Ejvind Mørtz, PhD
Co-Founder & COO, Alphalyse, Denmark
Ejvind holds a PhD from the University of Southern Denmark in Protein Chemistry & Molecular Biology. He has more than 20 years of experience developing protein analyses and mass spectrometry methods in the research & development of protein biologics; vaccines, monoclonal antibodies, therapeutic proteins, and cell and gene therapies.
LC-MS HCP assay validation and GMP release testing for complex samples
Thomas Kofoed, Co-Founder & CEO, Alphalyse, Denmark
30 May 2023 | Replay available below
Generic HCP-ELISAs often do not provide sufficient coverage of the Host Cell Proteins (HCPs) throughout the manufacturing process, thereby forcing you to develop a process-specific ELISA. However, the complexity of some products, like advanced therapies and proteins expressed in E. coli inclusion bodies, makes it impossible to produce suitable antigens for raising ELISA antibodies. Instead, many companies use Mass Spectrometry (MS) as an orthogonal method to ELISA – especially since MS can identify and even quantify impurities at an individual level.
Across the industry, it has been challenging to achieve LC-MS-based HCP analysis that complies with Good Manufacturing Practice (GMP) due to the complexity of its sample preparation and data analysis, the sophistication of high-end mass spectrometers, and the lack of reproducibility of the complete workflow.
This presentation gives an overview of the parameters that influence the reproducibility of mass spectrometry data, insights on how Alphalyse has optimized the technology to manage and track variability, shows a study of the reproducibility of mass spectrometry data across projects and time, and provides examples of client method qualification, included in their recent FDA approved IND.
Thomas Kofoed, PhD
Co-Founder & CEO, Alphalyse, Denmark
Thomas holds a PhD in Organic Chemistry and a BSc in Business Administration. He has 20+ years of management experience in the biotech industry and has developed protein analyses and mass spectrometry methods in the research & development of protein biologics.
HCP assays for pandemic vaccines: A case on adenovirus COVID-19 vaccines
Ejvind Mørtz, Co-Founder & COO, Alphalyse, Denmark
17 April 2023 | Replay available below
Preparedness against pandemic diseases demands rapid-response vaccine technology and ready-to-use analytical methods to support CMC activities. Manufacturing process consistency and comparability between GMP batches are important when the process is scaled up and the production is performed at several manufacturing facilities to provide a pure and safe product to vaccinate billions of healthy individuals.
Process-related impurity assays for pandemic vaccines should, therefore, ideally be a) very fast without long assay development time, b) generic for common manufacturing processes and vaccine types, and c) provide reproducible impurity measurements over time and in-between laboratories.
Here, we present results and learnings from LC-MS analysis of several viral-vector and protein-based Covid-19 vaccines on the market and under development. The results include the identity and quantity (ppm and ng/ml) of HCPs in marketed Covid-19 vaccines and a comparison of observed HCP profiles. We have found that the product purity and HCP profiles vary substantially between marketed Covid-19 vaccines: Some vaccines have almost a 1:1 ratio between viral proteins and HCPs, other vaccines having low ppm level impurities.
The presentation includes an assay performance evaluation regarding reproducibility and intermediate precession to support CMC manufacturing consistency and batch comparability.
Ejvind Mørtz, PhD
Co-Founder & COO, Alphalyse, Denmark
Ejvind holds a PhD from the University of Southern Denmark in Protein Chemistry & Molecular Biology. He has more than 20 years of experience developing protein analyses and mass spectrometry methods in the research & development of protein biologics; vaccines, monoclonal antibodies, therapeutic proteins, and cell and gene therapies.
Challenges and considerations in the HCP analysis for AAV-based gene therapy product
Yiling Bi, Sangamo Therapeutics, USA
28 February 2023 | LIVE ONLY
Host Cell Proteins (HCPs) are process-related impurities in a drug product that must be measured and continuously monitored due to potential safety concerns. Compared to traditional biologics, the quantitation and control of HCPs in gene therapy products face significant challenges as the manufacturing processes of these products are highly complex, with widely different upstream and downstream processes.
The HCPs have traditionally been measured by commercially available ELISA assays, which may or may not represent the specific process. In addition, HCPs contain both low and high-molecular-weight species, and it is important to get sufficient coverage to detect the majority if not all HCPs generated during the process. The HCP coverage can be measured using Antibody affinity extraction (AAE), 2D-Western analysis, and LC-MS-based approaches.
In this presentation, Yiling Bi will discuss a case study on the roadmap for HCP analysis for an AAV-based gene therapy product. In addition, some considerations for choosing (a) specific approach(es) during different clinical development stages will also be discussed.
Yiling Bi
Yiling Bi, Senior Scientist, Sangamo Therapeutics
Yiling Bi is a senior scientist in the analytical development department at Sangamo Therapeutics. Yiling is responsible for the characterization of AAV-based gene therapies using LC-MS and working with CRO/CTLs and vendors for routine testing and new technology evaluations.
LC-MS-based protein profiling of lentiviral vector products for CMC development
Albert Molina Gil, Orchard Therapeutics, UK
17 January 2023 | LIVE ONY
Lentiviral vectors used in the production of cell and gene therapy products present multiple sources of impurities derived from sources such as viral, starting materials, and process- and host-cell-derived constituents. While residual protein characterization for these products has typically been performed via conventional enzyme-linked immunosorbent assays (ELISAs), the robust identification and quantification of individual residual proteins from multiple sources require an appropriately matched analytical approach to ensure a solid and thorough characterization.
In this work, we have analyzed and compared residual protein levels of different lentiviral vector products across the various steps of the manufacturing process using liquid chromatography–mass spectrometry analysis. More than 200 media- and host-cell-related proteins were identified by two or more peptides, and >100 were quantified above the lower limit of quantification for intermediate samples across different products. Interestingly, therapeutic and viral proteins were also quantifiable, with the relative levels of the latter correlating with the expected stoichiometry. The analysis and respective data will be discussed in the context of Identification/quantification of the different vector components and process characterization and assessed as a tool to evaluate the consistency of lentiviral vector manufacturing platforms regarding protein profile.
Albert Molina Gil
Process Development Scientist, Orchard Therapeutics
Albert holds an industry-partnered PhD in Gene Therapy from University College London and GlaxoSmithKline. Following his PhD, he led the development of a stable cell line platform for gamma-retroviral vector production. Currently, he develops and optimizes platform technologies enabling the generation and characterization of stable cell lines and lentiviral vector production processes to treat genetic diseases.
About the event
Duration: 45 minutes, including Q&A
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