BEBPA Presents:

5th Annual Host Cell Protein Conference

May 10-12, 2017
San Francisco, CA USA

Speakers

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Roundtable Discussion Panel

  • Speaker: Svetlana Bergelson, Biogen
  • (Day 2: Roundtable Discussion)

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An Effective SWATH-MS Workflow for the Analysis of CHO HCPs

  • Speaker: Xuezhi Bi, Bioprocessing Technology Institute, A*STAR
  • Abstract: Residual host cell proteins (HCPs) impurity is one of the critical quality attributes (CQAs) in CHO-produced biotherapeutic monoclonal antibodies (mAbs). In order to monitor individual CHO HCP clearance, a sensitive, robust and effective SWATH-MS workflow was developed to identify and quantify these impurities. Using this workflow on premixed control samples comprising known ratios of pure mAb to residual HCPs, a total of 2413 proteins were identified with up to 60% of the proteins accurately quantified within ±20% of theoretical values. We next applied the workflow on downstream purification intermediates of a mAb product; we were able to identify 1300 and 1465 HCPs across the process intermediates in two independent runs, and to show significant reduction of 600 HCPs after Protein A affinity purification and two polishing steps. Subsequent spiking of the samples with known quantities of standard proteins enabled the absolute quantification of individual HCPs and resulted in improved detection sensitivity down to 1 fmol with a 1 µg total protein load – a five-fold higher sensitivity than the previously reported detection limit. To date, we have generated a comprehensive in-house CHO-K1 spectral ion library covering more than 8200 proteins (identified by ≥ 2 unique peptides at 1% FDR). Using a high performance streamlined software pipeline, we will demonstrate that the SWATH-MS workflow can be used for high-throughput and robust monitoring and quantification of HCPs during downstream purification processes of mAb production; the workflow also provides improved detection sensitivity and quantification accuracy of low-level HCPs.
  • ( Day 2: Utilizing Mass Spectrometry for Host Cell Protein Characterization Session)

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Performance of HCP LC-MS/MS scaled from nanoflow to standard LC flow using a Thermo Q Exactive Plus MS

  • Speaker: Jonas Borch-Jensen, Novo Nordisk
  • Abstract: Subject areas
     LC methods
     Scaling of LC methods
     Thermo MS platform
  • (Day 1: Workshop 1: LC-MS/MS for HCP Analysis)

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Anti-HCP Antibody Reagents and Assay Development From a CRO Perspective: Industry Wide Data, Trends and Recommendations

  • Speaker: Tobey Gooding, Covance
  • Abstract: (Pending)
  • (Day 1: Immunoassay Development & Integration with LC-MS)

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Overcoming the Challenges of Updating an HCP Control Strategy in a Marketed Product

  • Presenter: Feny Gunawan, Genentech
  • Abstract: In the lifecycle of a commercial product, updates to the manufacturing processes and control systems are to be expected. In the case of process related impurities, such as Host Cell Proteins (HCP), several unique challenges need to be considered when updating their control strategy. For any process change, the HCP should remain within the currently approved levels. However, as analytical technologies improved we are able to detect impurities at much lower levels and/or identify those that are previously unknown. This presentation will focus on a case study in which we updated the HCP control system of a marketed product, 20 years post approval, with modern analytical technologies and current regulatory expectations. In the midst of this update, a new co-purifying HCP — that was not detectable in the approved process — was discovered. The strategies and considerations for overcoming these challenges will be discussed.
  • Contributing Authors:
    Feny Gunawan1, Denise Krawitz3, Julie Nishihara2, Marty Vanderlaan3, and Heidi Zhang1
  • 1Analytical Operations, Genentech
  • 2Protein Analytical Chemistry, Genentech
  • 3Independent
  • (Day 2: Host Cell Protein Control Strategies for Pre- and Post-Marketed Products Session)

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CHO Strain vs. Culture Process: Comparing HCP Reagent Made From Different CHO Lines On Measurement of HCPs in the Same Products

  • Speaker: Lea Hagigi, Biogen
  • Abstract: (Pending)
  • (Day 1: Immunoassay Development & Integration with LC-MS Session)

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A holistic phase appropriate strategy for HCP-limits and HCP

  • Speaker: Markus Haindl, Biogen
  • Abstract: (Pending)
  • (Day 2: Host Cell Protein Control Strategies for Pre- and Post-Marketed Products Session)

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Quantitative Mass Spectrometry Strategies for Detection, Relative and Absolute Quantitation of Host Cell Proteins. Data dependent, Independent and targeted analyses

  • Speaker: Eric B. Johansen, AbbVie
  • Abstract: Quantitative mass spectrometry strategies are rapidly developing greater sensitivity and selectivity. Here we will discuss workflows and best practices from detection and relative abundance to absolute quantitation by LCMS as they relate to host cell proteins and peptides.
  • (Day 1: Workshop 1: LC-MS/MS for HCP Analysis)

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Identification and Quantification of HCP’s by SWATH LC MS for Process Development of a Novel Biologic

  • Speakers: Thomas Kofoed, Alphalyse, and Lars Skriver, Savara Pharmaceuticals
  • Abstract: HCP analysis by ELISA for novel biologics under development is difficult because no platform or product specific ELISA has been developed yet. For biologics expressed in E.coli inclusion bodies it may not even be possible to develop a product specific ELISA because immunization reagents also contain the drug substance.
    Here we present a HCP analysis strategy and analysis results for a novel biologic, where a platform ELISA is not available, using generic ELISA methods in combination with SWATH LC MS/MS for identification and quantification of individual HCP’s.
    The generic ELISA assays are based on off-the-shelf kits from different vendors. The SWATH LC MS/MS method is based on a 1D microflow LC method in combination with SWATH data independent acquisition on a Sciex trippleTOF 6600 mass spectrometer. The microflow 1-dimensional chromatography enables very fast and robust chromatography compared to multi-dimensional nanoflow chromatography. The SWATH independent data acquisition enables very reproducible HCP identification and quantification compared to data dependent workflows.
  • Contributing Authors:
    Thomas Kofoed, Marie Grimstrup, Janne Crawford, Jakob Bunkeborg, Ejvind Mørtz, Alphalyse A/S, Denmark.
    Lars Skriver, René Egebro, Savara Pharmaceuticals Austin, TX, USA.
  • (Day 1: Host Cell Protein Impurities & Product Quality Session)

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Workshop 2 Instructor

  • Speaker: Denise Krawitz, Consultant
  • (Day 1: Workshop 2: Host Cell Proteins 101)

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Roundtable Discussion Panel

  • Speaker: Denise Krawitz, Consultant
  • (Day 2: Roundtable Discussion)

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The 2017 Chinese Hamster Reference Genome

  • Speaker: Kelvin Lee, University of Delaware
  • Abstract: (Pending)
  • (Day 2: Utilizing Mass Spectrometry for Host Cell Protein Characterization Session)

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Case Study for a Validation of a MS Based HCP Quantification Method – Requirements and Limitation

  • Speaker: Ingo Lindner, Roche
  • Abstract: (Pending)
  • (Day 2: Utilizing Mass Spectrometry for Host Cell Protein Characterization Session)

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Case Study: Clinical perspectives on Phospholipase B-Like 2 Protein, a Host Cell Impurity in Lebrikizumab Clinical Material

  • Speaker: John Matthews, Genentech
  • Abstract: (Pending)
  • (Day 1: Host Cell Protein Impurities & Product Quality Session)

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Lessons Learned: HCP Analysis of a Variety of Biologics and Biosimilars using LC-MS/MS

  • Speaker: Laura McIntosh, Caprion Biosciences
  • Abstract: (Pending)
  • (Day 1: Host Cell Protein Impurities & Product Quality Session)

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Case Study: HCP Antigen Stability

  • Speaker: Emily Menesale, Biogen
  • Abstract: (Pending)
  • (Day 1: Immunoassay Development & Integration with LC-MS Session)

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Roundtable Discussion Panel

  • Speaker: Ned Mozier, Pfizer
  • (Day 2: Roundtable Discussion)

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HCP Control Strategy Reassessment for a CHO-derived Protein Therapeutic

  • Speaker: John Rolf, Amgen
  • Abstract: (Pending)
  • (Day 2: Host Cell Protein Control Strategies for Pre- and Post-Marketed Products Session)

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Data Analysis, Quantitation and Reporting for Mass Spectrometry-Based HCP Studies

  • Speaker: Oliver Schirm, Caprion Biosciences Inc.
  • Abstract:The high specificity and sensitivity of mass spectrometry enables high HCP coverage, providing identification and quantitative information on individual HCPs. Non-targeted LC-MS/MS based workflows and targeted LC-MRM/MS workflows have been developed which routinely achieve sensitivity in the 1-10ppm range for both HCP identification and absolute quantitation. Case studies will be shown which illustrate how data analysis, quantitation and reporting can be performed for various HCP applications, including, for example: characterization of in-process samples and DS, comparability studies of Biosimilars to Innovators and absolute quantification of HCPs. For non-targeted data analysis, creation of appropriate databases for CHO, E.coli and human production systems will be covered, in addition to selection of false discovery rates, usage of different search engines and spectral counting vs. intensity based measurements. For quantitative data analysis, an automated adaptation of Skyline will be presented.
  • Contributing Authors:
    Oliver Gingras, Laetitia Cortes, Laura McIntosh, Michael Schirm
  • (Day 1: Workshop 1: LC-MS/MS for HCP Analysis)

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Monitoring & Identifying Critical HCPs During CHO Cell Bioprocessing: Do They Exist & Why We Shouldn’t Have to Reclibrate HCP Assays Between Projects

  • Speaker: Mark Smales, University of Kent
  • Abstract: (Pending)
  • (Day 1: Host Cell Protein Impurities & Product Quality Session)

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Increased Throughput and Accuracy in Host Cell Protein Quantitation using Spectral Library Searches

  • Speaker: Martha Stapels, Sanofi Genzyme
  • Abstract: (Pending)
  • (Day 1: Immunoassay Development & Integration with LC-MS Session)

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Workshop 1 Instructor

  • Speaker: Kevin Van Cott, University of Nebraska
  • (Day 1: Workshop 2: Host Cell Proteins 101)

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Host Cell Proteins Seen as Critical Quality Attributes

  • Speaker: Thomas Waerner, Boehringer Ingelheim
  • Abstract: (Pending)
  • (Day 1: Host Cell Protein Impurities & Product Quality Session)

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Roundtable Discussion Panel

  • Speaker: Thomas Waerner, Boehringer Ingelheim
  • (Day 2: Roundtable Discussion)

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Knowledge Based HCP Risk Control Strategy During Downstream Process Development with Recent Case Studies

  • Speaker: Fengqiang Wang, Merck
  • Abstract: (Pending)
  • (Day 1: Host Cell Protein Impurities & Product Quality Session)

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Roundtable Discussion Panel

  • Speaker: Michael Wiedman, Boehringer Ingelheim
  • (Day 2: Roundtable Discussion)

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Development of a Fast LC-MS/SWATH Method for Real-Time Identification and Quantitation of the Residual HCPs During the Manufacturing Process of a Monoclonal Antibody Therapeutic

  • Speaker: Boyan Zhang, Mab-Works
  • Abstract: Subject areas
     DIA – SWATH
     Quantitation
     Case-study, real system
  • (Day 1: Workshop 1: LC-MS/MS for HCP Analysis)

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Quantitative Investigation of HCP Impurities: Bridging the Gap Between ELISA & Orthogonal LC-MS/MS Analysis

  • Speaker: Ying Zhang, Pfizer
  • Abstract: (Pending)
  • (Day 1: Immunoassay Development & Integration with LC-MS Session)