Cambridge Healthtech Institute’s Inaugural
Host Cell Protein: Detection, Analysis and Removal
Detecting, Analysing and Removing Host Cell Proteins
Part of CHI's 8th Annual The Bioprocessing Summit
August 15-16, 2016 | Westin Boston Waterfront | Boston, MA
The presence of residual Host Cell Proteins is generally considered a critical quality attribute due to the impact on product safety and efficacy. Assay-based methods are available but limited in their coverage and specificity, with questions still remaining
about the role of new analytical technologies such as mass spectrometry.
CHI’s inaugural Host Cell Proteins conference brings together the biotech ecosystem to discuss the latest challenges in HCP analysis, risk assessment and control strategies,
assay coverage, critical reagents and platforming, plus questions relating to HCPs testing in biosimilars, in-process testing and the latest data supporting the link between HCPs and immunogenicity.
Final Agenda
Monday, August 15
8:00 am Short Course Registration
9:00 – 11:30 Recommended Morning Short Course*
SC3: New USP Initiatives and Standards for Biologics
* Separate registration required
11:30 Main Conference Registration Opens
1:00 pm Chairperson’s Opening Remarks
Judy Shimoni, Ph.D., Senior Scientist and Group Leader, PTDU, Protein Analytical Chemistry, Genentech
1:10 KEYNOTE PRESENTATION:
Best Practices for Host Cell Protein Measurement: USP Chapter <1132>
Maura Kibbey, Ph.D., Director, Science and Standards, Global Biologics, USP
USP’s official chapter <1132> provides expert guidance on (1) HCP immunogen and standard preparation and characterization; (2) immunization; (3) purification and characterization of antibodies; (4) immunoassay development and validation;
(5) lifecycle management; and (6) orthogonal tools to supplement the primary immunoassay technology. Due to these complex, multi-antigen HCP immunoassays, emphasis is placed on unique challenges (e.g., sample dilution linearity, validation, and
reporting of results).
1:45 HCP Testing, Control Strategy and Risk Assessment
Judy Shimoni, Ph.D., Senior Scientist and Group Leader, PTDU, Protein Analytical Chemistry, Genentech
The presentation is intended to share some current thinking and to use case studies to demonstrate the application of orthogonal methods complementing HCP workhorse testing method and supporting process development and control strategy; also to illustrate
the use of risk assessment for HCP identification and control with consideration of clinical information.
2:15 Immunogenicity Risk Assessment of Host Cell Protein in Therapeutic Monoclonal Antibodies
Qingchun Zhang, Ph.D., Senior Scientist, Amgen
In this study, we investigated the HCP profiles for partially purified in-process samples and highly purified DS-like samples for four therapeutic mAbs. The results indicated in-process samples with up to 4000 ppm HCP content (levels >200 times greater
than the drug substance) did not pose a higher immunogenicity risk than highly-purified DS samples. Furthermore, sequence associated immunogenicity risk of over 20 common HCPs was evaluated using in silico algorithms.
2:45 Refreshment Break
3:15 HCP Assays as In-Process Tests
Carl
Co, Ph.D., Senior Scientist, Biogen Idec
HCPs must be removed to ensure product efficacy and safety. ICH specifications guidance, Q6B, states that for certain impurities, testing of drug substance may not be included in specifications if control or removal to acceptable levels is demonstrated.
Case studies will be presented which highlight the challenges of our strategy of performing HCP assays for in-process and not for release.
3:45 Sponsored Presentation (Opportunity Available)
4:00 Platform Reagents. The Design-Space of Generating Platform Reagents for HCP Monitoring by Assessing the Impact of Upstream Process Condition Variations on HCP Expression Profile
Fengqiang Wang, Ph.D., Associate Principal Scientist, Merck Research Laboratories, Merck & Co. Inc.
Mock host cell proteins produced from a null cell line without the product-encoding gene are commonly used as the calibration standards for HCP ELISA. The generation of mock HCPs often follows similar or identical upstream manufacturing process as the
production cell culture. We explored the upstream process range or design space where a platform assay can remain to be relevant and suitable for monitoring HCP clearance.
4:30 Breakout Discussions
This session provides an opportunity to discuss a focused topic with peers from around the world in an open, collegial setting. Select from the list of topics available and join the moderated discussion to share ideas, gain insights, establish collaborations, or commiserate about persistent challenges.
USP Feedback on HCPs
Moderator:
Maura Kibbey, Ph.D., Director, Science and Standards, Global Biologics, USP
Oliver Anderka, Ph.D., Fellow, Functional Lead Bioanalytics, Novartis Pharma AG
Risk Assessment Strategies for HCPs
Moderator:
Judy Shimoni, Ph.D., Senior Scientist and Group Leader, PTDU, Protein Analytical Chemistry, Genentech
5:30 Grand Opening Reception in the Exhibit Hall with Poster Viewing
7:00 End of Day
Tuesday, August 16
7:30 am Registration Opens and Morning Coffee
7:55 Chairperson’s Remarks
Phoebe Baldus, Ph.D., Principal Scientist and Group Leader, Pfizer
8:00 Adding MS to a Host Cell Protein Workflow
Phoebe Baldus, Ph.D., Principal Scientist and Group Leader, Pfizer
8:30 Analysis of Host Cell Proteins throughout Biopharmaceutical Purification
Martha Stapels, Ph.D., Senior Scientist, Biopharmaceuticals Development, Sanofi Genzyme Corporation
Methods have been developed to identify and quantify HCPs by mass spectrometry, but these typically involve 2D chromatography. We have developed a high throughput method to analyze HCPs where peptide identification occurs where the HCP is most
abundant, followed by quantitation in samples across the purification process. Statistical analysis is then used to evaluate purification steps and optimize the process.
9:00 Utilization of Mass Spectrometry for HCP Analysis during Process Development
Gang Xiao, Ph.D., Scientist, Process Development, Amgen
It is critical to measure individual HCP by mass spectrometry in order to troubleshoot or optimize the bioprocess. In this presentation, we will demonstrate how mass spectrometry can be utilized for HCP analysis during the bioprocess from upstream
cell production, through downstream purification, to final product releasing.
9:30 HCP Identification and Absolute Quantification with a Multifaceted Mass Spectrometry Platform
Laura McIntosh, Director, Vice President, Translational Research, ProteoCarta Management, CAPRION
A highly sensitive discovery platform has been developed, employing fractionation, deglycosylation, custom databases, and multiple search engines, to enable the identification and relative quantification of low ppm HCP from any host cell (CHO,
human, yeast, bacteria), without the need to remove the drug substance. Targeted MRM mass spectrometry is then used for absolute quantification down to 1 ppm. These assays enable process improvement, clearance monitoring, determination
of reproducibility, impact of scale-up and other monitoring needs for antibodies, blood products or peptide manufacture.
9:45 Coffee Break in the Exhibit Hall with Poster Viewing
10:30 Quantitative MS/MS in the Determination of HCP Levels
Nesredin A Mussa, Ph.D., Head, Process Analytics and Method Development, Biologics Development, Global Manufacturing and Supply, Bristol-Myers Squibb
11:00 Host Cell Protein Analysis by Mass Spectrometry and Its Application in a Comparability Exercise
Michael Gattner, Ph.D., Associate
Scientist, Physico-Chemical Characterization, Sandoz Biopharmaceuticals/ Hexal
Host cell proteins (HCPs) are process-related impurities present in biopharmaceuticals and are generally considered to be critical quality attributes. Here we discuss the usage of ELISA and mass spectrometry to monitor HCP populations
in the context of comparability and similarity excercises. A case study employing mass spectrometry-based HCP analysis following a manufacturing change is provided.
11:30 Host Cell Proteins: The Hidden Side of Biosimilarity Assessment
Roxana Butoi, Ph.D., Manager, Analytical Development, R&D Platform Technology and Science, GlaxoSmithKline
Results illustrate the efforts to assess “biosimilarity” for a specific group of process-related impurities, the host cell proteins (HCP). Extensive characterization of HCP in the drug substance of a biosimilar candidate guided
process development to improve HCP clearance. The data presented illustrate the challenge of matching the reference product on either quantitative or qualitative aspects of HCP impurities.
12:00 pm Integration of Data from Orthogonal Methods to Provide a Comprehensive Analysis of HCPs in Final Drug Substance
Kenneth Hoffman, Ph.D., President, Cygnus Technologies
Because of certain limitations of ELISA, orthogonal methods of analysis are required. This talk discusses how data from 2D HPLC, Antibody Affinity Extraction (AAE), 2D PAGE, and LCMS/MS when integrated with ELISA data can be used to
give a comprehensive analysis of individual HCPs that persist through a purification process.
12:30
Luncheon Presentation: Reducing Operator Intervention with a Comprehensive Approach to HCP and Residual Protein A Testing
Renee Tobias, Product Manager, Pall ForteBio LLC
Using an automatable alternative to ELISAs for Host Cell Protein and Residual Protein A detection could significantly reduce the number of manual assay steps – and the variability those steps introduce. The Ready BLI kits from
Pall ForteBio provide a more complete solution to impurity testing in bioprocess development and in-process QC.
1:15 Dessert Refreshment Break in the Exhibit Hall with Poster Viewing
1:55 Chairperson’s Remarks
Fengqiang Wang, Ph.D., Associate Principal Scientist, Merck Research Laboratories, Merck & Co. Inc.
2:00 Platform Host Cell Protein Assays: How to Demonstrate Fitness for Use
Oliver Anderka, Ph.D., Fellow, Functional Lead Bioanalytics, Novartis Pharma AG
“Platform” host cell protein assays are used by biomanufacturers to test a variety of products made from the same type of host organism. An outline is given of how to demonstrate a platform assay’s fitness for use. This includes reagent characterization/qualification, reagent replacement, assay validation, as well as criteria for application of a platform assay to a new product or process.
2:30 HCP Analysis for a Spectrum of Products Generated from Multiple CHO Cell Lines Including a Comparison of the Performance of a Panel of Different HCP Commercial Kits
Girija Krishnamurthy, Ph.D., Director, Method Development, Bristol-Myers Squibb
3:00 Cover Your Assay: Approaches to Evaluate HCP ELISA Reagent Coverage
Denise Krawitz, Ph.D., Senior Manager Analytical Operations, Genentech
In order to use ELISAs, we must demonstrate that the antibodies used in the assay recognize a majority of the proteins produced by the host cell. While 2D SDS-PAGE and Western blots techniques are valuable for a qualitative assessment of antibody coverage, they have limited value as a quantitative tool. Additionally, platform HCP ELISAs are used to monitor HCPs in multiple products. Proteomics studies of different cell lines grown under different culture conditions suggest that the vast majority of proteins expressed are the same between cell lines.
3:30 Refreshment Break in the Exhibit Hall with Poster Viewing
4:15 Finding Needles in a Haystack – Strategies and Tools for HCP Identification In Biopharmaceuticals
Xiaohui Lu, Ph.D., Senior Scientist, BioPharma Development, Biogen
Identification of host cell proteins (HCPs) in biopharmaceuticals drug is hampered by the presence of overwhelming amount of the products. This case study compared different strategies to remove product and enrich HCPs. Applying these
enrichment technologies will greatly improve MS-based HCP identification, and provide much needed HCP information for purification process development and risk assessments.
4:45 High Throughput HCP Assays for Bioprocess Development
Christopher Larkin, Ph.D., Scientist II, MedImmune a Member of the AstraZeneca Group
Purification processes without an affinity capture step often require extensive process development, potentially generating many thousands of samples for HCP testing. Analytical approaches to meet the required throughput, whilst producing
high quality data, are desirable. Here, we describe the use of a high throughput platform for HCP testing to enable rapid process development and optimization. Case studies will be presented where we have employed this approach
to successfully reduce and control HCP levels.
5:15 End of Conference
5:15 Registration for Dinner Short Course