Cambridge Healthtech Institute’s 3rd Annual

Host Cell Proteins

Detection, Analysis and Control

August 13-14, 2018

A critical part of bioprocessing is the control of process-related impurities such as host cell proteins (HCPs), which co-purify with the drug substance and can cause adverse effects such as immunogenicity. Analytical methods are available but coverage and specificity is limited. Moreover, the emergence of new expression systems, new products and new techniques such as mass spectrometry has further compounded these limitations with regulators now pressing companies for more HCP data.

CHI’s Host Cell Proteins conference brings together industry leaders to discuss critical HCP topics such as: risk assessment and control strategies, HCP characterization, assay coverage, critical reagents and platforming, plus questions relating to biosimilars, in-process testing and the latest data supporting the link between HCPs and immunogenicity.

Final Agenda

Monday, August 13

8:00 am Short Course Registration Open (Grand Ballroom Foyer) and Morning Coffee (Break Foyer)

9:00 - 11:30 Recommended Short Course*

SC5A: Introduction to Host Cell Proteins: Analysis, Characterization, Risks, and Requirements

Instructor: Denise Krawitz, PhD, Principal Consultant, CMC Paradigms LLC

* Separate registration required.

11:30 Main Conference Registration Open (Grand Ballroom Foyer)

HCP REGULATIONS AND STANDARDS
Back Bay B

1:00 pm Chairperson’s Opening Remarks

Denise Krawitz, PhD, Principal Consultant, CMC Paradigms LLC

1:10 KEYNOTE PRESENTATION: New USP Initiatives to Support Host Cell Protein Analytical Methods

Maura Kibbey, PhD, Director, Global Biologics, U.S. Pharmacopeia

Impurities such as host cell proteins (HCPs) must be cleared from recombinant biotherapeutics to minimize safety concerns and ensure the quality of the drug substance. HCP ELISA’s can miss specific proteins that may persist, and therefore the use of orthogonal methods has increased. USP’s HCP Expert Panel developed General Chapter <1132> to describe best practices for measurement of HCPs. This presentation will include an update on USP standards to support characterization and measurement of specific HCPs.

1:45 FEATURED PRESENTATION: Regulating Host Cell Proteins

Erika Friedl, PhD, Quality Expert, Hematology and Transfusion Medicine, Paul-Ehrlich-Institute

Specific HCP regulatory guidance is outlined in the European Pharmacopoeia and the US Pharmacopoeia, which provide an important regulatory framework. The interpretation of the regulatory framework and its flexibilities will be discussed. Case studies will be presented to outline the expectations regarding the HCP control strategy, the specification setting and the type of HCP assays used during product development and in the licensing process.

2:15 Use of Host Cell Protein Risk Assessments to Help Ensure Patient Safety, Product Quality, and Manufacturing Process Consistency

Denise Krawitz, PhD, Principal Consultant, CMC Paradigms LLC

With improvements in analytical technologies, such as mass spectrometry, identification of individual HCP impurities has become more commonplace during clinical development over the past several years. As we learn more about HCP impurities, we can better understand the impact HCPs can have on the stability and safety of biotechnology products. This presentation will cover HCP risk assessment strategies to help ensure patient safety and product quality throughout product development.

2:45 Refreshment Break (Foyer)

HCP ASSAY DEVELOPMENT
Back Bay B

3:15 Same but Different: A Case Study of HCP Comparability for an In-Licensed Project

Feny Gunawan, PhD, Scientist, Analytical Operations, Genentech

This presentation will focus on a case study in which we evaluated the HCP comparability of an in-licensed project. The challenges that we were facing were two-fold: 1) to determine the HCP comparability of the in the in-house produced material to the previous company’s material that was used in the pivotal trial, 2) to assess the suitability of HCP ELISA – a commercial HCP ELISA was compared to our platform HCP ELISA.

3:45 The Journey to Replace an Existing HCP Assay - Starting with the Right Antigen

Emily Menesale, Senior Associate Scientist, Analytical Development, Biogen

Measuring residual HCP is essential for the bioproduction process. HCP assay results are reagent-dependent by nature, so there are significant challenges when it comes to replacing existing HCP reagents. Here, we describe the careful selections taken in our antigen processing and characterization to ensure an optimized antigen prep. This antigen ultimately led to antibody production and a successful HCP assay development that resulted in a suitable HCP replacement method.

4:15 Common Mistakes and Myths of HCP Analysis - Integration of Orthogonal Methods to Detect Individual Downstream HCPs

Ken Hoffman, President, Cygnus Technologies

Various regulatory guidance articulate better practices in HCP analytics. However, the repertoire of HCP analytics still contains several outdated conventional practices. This talk identifies several mistakes and myths regarding HCP analytics, discusses proper methods to generate HCP antibodies and develop ELISAs. With Antibody Affinity Extraction (AAE) to replace 2D WB, and improvements in Mass Spectrometry, the identification of individual downstream HCPs is now within our capability and should be the goal of any well-characterized biologic.

5:30 Grand Opening Reception in the Exhibit Hall with Poster Viewing (Grand Ballroom)

7:00 End of Day

Tuesday, August 14

7:30 am Registration Open and Morning Coffee (Grand Ballroom Foyer)

IMPROVING HCP ANALYSIS WITH MASS SPEC
Back Bay B 

7:55 Chairperson’s Remarks

Yan-Hui Liu, PhD, Merck & Co., Inc.

8:00 Making the Most of Mass Spectrometry in Host Cell Protein Detection and Control

Christopher Yu, PhD, Principal Scientist, Genentech, A Member of the Roche Group

Identification of specific residual host cell proteins (HCP) in biopharmaceutical products by mass spectrometry addresses potential gaps in immunodetection coverage. This study examines the technical aspects of HCP quantitation by mass spectrometry, and demonstrates feasibility of fast and relevant quantitation for optimal process development. We also discuss the importance and potential opportunities in defining the quantitative ranges of HCP for safe and efficacious use of biopharmaceuticals.

8:30 Qualification of an Absolute Quantification Workflow for HCP Determination by LCMS

Veronika Reisinger, PhD, Lab Head, Biologics Technical Development and Manufacturing, Novartis

ELISA is the standard method for HCP detection but is usually not capable to identify and quantify single HCPs. For tracking of a specific HCP, an absolute quantification workflow based on LCMS/MS was developed and qualified regarding different parameters like linearity or repeatability. The method was applied to support process optimization as well as increase knowledge on the final drug substance.

9:00 Determination of Proteins Detected by Host Cell Protein ELISA Assays Using Mass Spectrometry

Michelle Busch, PhD, Scientist, Bioanalytics Characterization, Sanofi

Mass spectrometry is typically used as an orthogonal HCP characterization technique while an ELISA method is used as the release assay. Sometimes, ELISA assays have been shown to be insensitive to relatively abundant HCPs. In this study, the antigens used to raise ELISA antibodies were analyzed to measure amounts of HCPs and then the antibodies were used to purify HCPs from products to better understand ELISA assay results.

9:30 Summary and Feedback on Roundtables 

9:45 Coffee Break in the Exhibit Hall with Poster Viewing (Grand Ballroom)

OPTIMIZING HCP ANALYSIS
Back Bay B

10:30 Tracking Host Cell Protein Impurities in Biopharmaceuticals Using Mass Spectrometry

Regina Kufer, PhD, Senior Associate Development Analytics, Roche Diagnostics GmbH

Recently a MS-based platform was developed for host cell protein (HCP) characterization to support purification process development. This technique provides an orthogonal and complementary approach to traditional HCP analysis by enzyme-linked immunosorbent assays (ELISA). In contrast to the ELISA, LC-MS/MS analysis provides the identity of HCPs. Here, we present a simple and powerful strategy combining our platform LC-MS/MS with a proteomic approach to increase sensitivity and number of detected HCPs.

11:00 Advances in HCP Analysis for the Quality Control Laboratory

Jamie C. Rusconi, PhD., Staff Analytical Scientist, Bioanalytical Development, Regeneron

While ELISA assays generally offer the advantages of broad HCP recognition and sensitivity, they also suffer from a limited linear dynamic range and multiple steps requiring analyst “hands on” time. The ELLA instrument from ProteinSimple has been developed as an alternative to the traditional immunoassay format. In this presentation, we will explore the application of the ELLA instrument from ProteinSimple as an alternate approach to ELISA for HCP analysis.

11:30 High Throughput LC-MS Platform for HCP Characterization in Support of Process Development

Suli Liu, PhD, Scientist II, Biogen

As an orthogonal method to ELISA, LC-MS based analytical approach offers unique features of identification and quantitation of individual HCPs. A high throughput, high sensitivity and robust LC-MS based platform has been developed to support HCP clearance monitoring during process development. This platform relies on 1D LC coupled with advanced mass spectrometry and combined with ‘Hi 3’ quantitation method achieves detection and quantitation of HCPs in a single run. The method has been optimized for up to 19 samples per day with low ppm detection limit. This presentation will discuss the method development, qualification and a case study.

12:00 pm Characterization, Optimization and Application of an LC-MS Based Workflow for HCP Identification and Control

Michael Schirm, PhD, Associate Director, Research & Development Proteomics, Caprion Biosciences, Inc.

Mass spectrometry (MS) enables identification and quantitation of total and individual HCP in biotherapeutic products, and represents an orthogonal method to ELISA. Examples will be presented showing use of semi-quantitative identification of individual/total HCP (LC-MS/MS) and absolute quantitation of HCP (LC-MRM/MS), as applied to monitoring of process changes/improvements, scale-up, batch uniformity, clearance, and comparison of Biosimilars vs. Innovators. Caprion’s HCP platform features customizable organism/process-specific databases, highly controlled analytical processes and reproducible robust detection (to ~1ppm).

12:30 Luncheon Presentation: Host Cell Proteins-Integrated Analytical Solutions Using Immunoassays and Mass Spectrometry

Andrew Hanneman, PhD, Associate Director, Biologics Testing Solutions, Charles River Laboratories

Proteomic mass spectrometry methods have been used to identify and quantitate specific HCPs in biotherapeutics, and their process removal into drug substance. The protein-specific identification potential of MS vs. the high throughput and ease-of-use of ELISAs has made it challenging to unite the two strategies. Protein pull-down methods can be used to leverage the power of MS to characterize ELISA reagent HCP coverage offering a complementary methodology to identify and and target HCPs throughout bioprocessing.

1:15 Dessert Refreshment Break in the Exhibit Hall with Poster Viewing (Grand Ballroom)

HOW HCP ANALYSIS IMPACTS BIOPROCESSING
Back Bay B

1:55 Chairperson’s Remarks

Yan-Hui Liu, PhD, Merck & Co., Inc.

2:00 Evaluation of Analytical Technology to Accelerate Process Development and Support Next-Generation Manufacturing

Caitlin Kramer, PhD, Associate Scientist, Process Development Analytics, Bristol-Myers Squibb

Instruments were evaluated for their potential application in substituting for traditional ELISA impurities assays. The options were evaluated from both a high-throughput process development perspective and a process analytical technology perspective, for future use in next-generation manufacturing scenarios.

2:30 Identification and Impact of Individual Host-Cell Proteins in CHO-Based Biomanufacturing

Abraham M. Lenhoff, Allan P. Colburn Professor, Department of Chemical and Biomolecular Engineering, University of Delaware

Overall levels of host-cell proteins (HCPs) in biomanufacturing are routinely monitored by methods such as ELISA. However, even when such levels are reduced to acceptable levels, individual HCPs may be problematic within a bioprocess or may impact drug product quality. This presentation will discuss the identification of such problematic HCPs, possible reasons for their persistence, and remedial actions that may be employed to address the deleterious effects.

3:00 An Improved CHO HCP Detection System for Biopharmaceuticals

Mindy Wan, PhD, Technical Director, Business Development, KBI Biopharma, Inc.

KBI has applied a unique strategy to develop an improved CHO HCP detection system for a variety of protein therapeutics expressed in CHO cell lines widely used in biopharmaceutical industry. KBI’s CHOP ELISA method exhibits superior dilution linearity and matrix flexibility for detecting CHOPs present in purification process intermediates and final products. The reagent used in this ELISA shows >70% coverage for a pooled null harvest generated from several CHO cell lineages.  

 3:30 Refreshment Break in the Exhibit Hall with 1st Session Poster Winner Announced (Grand Ballroom)

4:15 Linking HCPs to Bioprocess Performance

Ying Zhang, PhD, Senior Scientist, Analytical Research & Development, Pfizer

LC-MS/MS has emerged as an orthogonal approach to ELISA for analyzing HCPs, providing both qualitative and quantitative information on individual HCPs. This presentation will discuss the use of LC-MS/MS to improve the understanding of bioprocessing.

4:45 HCPs Analysis by Mass Spectrometry for Bioprocess Development

Yi Wang, PhD, Senior Scientist, Protein Mass Spectrometry Department, Merck & Co., Inc.

CHO host cell proteins (HCPs) are process-related impurities that need to be monitored during process development and drug substance release of biologic therapeutics, due to their potential impacts on drug safety, drug stability and drug efficacy. Although ELISA is the industry gold standard for residual HCP detection, MS-based methods have been developed for HCP identification and relative quantitation due to its superior selectivity and sensitivity. In this talk, several case studies will be presented to demonstrate how different MS strategies are used for bioprocess development.

5:15 End of Conference