Advanced Host Cell Protein (HCP) Analysis Training Course

Advanced Host Cell Protein (HCP) Analysis Training Course

Introduction

Host Cell Proteins are a critical class of process-related impurities in the manufacture of bio therapeutics, including monoclonal antibodies, recombinant proteins, vaccines, and gene therapies. Their presence, even at parts-per-million levels, can compromise a drug's safety, stability, and efficacy, potentially causing adverse immune responses or impacting the final product's critical quality attributes. Consequently, rigorous and state-of-the-art HCP analysis is non-negotiable for regulatory compliance. The industry is rapidly moving beyond reliance on traditional generic ELISA methods, which face limitations in terms of antibody coverage and identification of low-abundance or high-risk HCPs. Advanced Host Cell Protein (HCP) Analysis Training Course is designed to bridge the gap between conventional approaches and the sophisticated demands of modern bio manufacturing..

The future of bioprocess development and quality control (QC) lies in integrating orthogonal analytical methods, particularly Liquid Chromatography-Mass Spectrometry (LC-MS), to achieve deep-profiling of the HCP complement. Participants will gain mastery over advanced techniques such as SWATH-MS and Multiple Reaction Monitoring (MRM) for specific HCP quantification and identification. Crucially, the course emphasizes a Quality by Design (QbD) and risk-based control strategy approach, enabling scientists to track problematic HCPs throughout downstream processing (DSP). By mastering these advanced methodologies, professionals will be equipped to secure GMP batch release, accelerate time-to-market, and ensure the highest standards of patient safety in the burgeoning global biopharmaceutical market.

Course Duration

10 days

Course Objectives

1. Master the principles of LC-MS/MS for HCP identification and quantification, positioning it as the primary orthogonal method.
2. Evaluate and apply risk-based control strategies for managing Critical Quality Attributes (CQAs) related to HCPs.
3. Develop robust, process-specific ELISA protocols and conduct in-depth antibody coverage analysis
4. Interpret and comply with evolving regulatory guidance from the FDA, EMA, and ICH on residual impurities.
5. Implement advanced Mass Spectrometry (MS) techniques, including SWATH-MS and MRM, for high-sensitivity, multiplexed HCP monitoring.
6. Design and execute effective HCP clearance studies to inform and optimize downstream processing (DSP) steps.
7. Identify and track Problematic HCPs with high-risk potential for product instability or immunogenicity.
8. Apply proteomics and bioinformatics tools for in-depth HCP deep-profiling and data interpretation.
9. Integrate Quality by Design (QbD) principles into the entire HCP control strategy lifecycle.
10. Troubleshoot common issues in both immunoassay and MS-based analytical workflows.
11. Compare and select the most fit-for-purpose analytical platform based on product development phase
12. Establish scientifically justified acceptance criteria and HCP limits in the final drug substance
13. Ensure data integrity and prepare comprehensive documentation for successful regulatory submissions.

Target Audience

1. Analytical Development Scientists
2. Bioprocess/Downstream Processing (DSP) Engineers
3. Quality Control (QC) Analysts and Managers
4. Regulatory Affairs Professionals
5. Formulation and Stability Scientists
6. Principal Investigators/Project Leaders in Biologics R&D
7. Quality Assurance (QA) Specialists
8. Contract Research/Manufacturing Organization (CRO/CDMO) Personnel

Course Modules

Module 1: HCPs as Critical Quality Attributes (CQA) in Biologics

• Definition, classification, and source of Host Cell Proteins
• How HCPs cause immunogenicity and product instability.
• Regulatory landscape: ICH Q6B and phase-appropriate HCP limits
• HCP expression dynamics across different bioreactor and harvest conditions.
• Case Study: Analysis of an approved mAb demonstrating the impact of a residual HCP on Fc region stability.

Module 2: Fundamentals of HCP Immunoassay (ELISA) Development

• Pros, cons, and selection criteria.
• Strategies for anti-HCP antibody generation and effective host cell immunization
• Assay qualification and validation for specificity, sensitivity, and precision.
• Hook effect, lot-to-lot variability, and matrix effects.
• Case Study: Troubleshooting a commercial ELISA failure due to low signal in a high-concentration drug substance matrix.

Module 3: Antibody Coverage Analysis and Evaluation

• 2D-PAGE, Western Blot, and 2D-DIGE for antibody reactivity.
• Quantitative assessment of HCP antibody coverage
• Statistical metrics for defining acceptable coverage in process-specific assays.
• Orthogonal method integration to confirm ELISA performance (LC-MS/MS).
• Case Study: Using 2D Western Blot to demonstrate lack of coverage for a major p-HCP co-purifying with the product.

Module 4: Introduction to Mass Spectrometry (MS) as an Orthogonal Tool

• Ionization (ESI) and mass analysis (Orbitrap, TOF) for protein identification.
• HCP digestion (trypsin) and peptide mapping.
• LC-MS/MS system configuration and data-dependent acquisition (DDA).
• Advantages of MS: Individual HCP identification and independence from antibodies.
• Case Study: Using MS data to resolve a discrepant ELISA result from a new DSP iteration.

Module 5: Advanced LC-MS for HCP Quantification: SWATH & DIA

• Concept of Data-Independent Acquisition (DIA) vs. DDA for comprehensive data collection.
• Mastering SWATH-MS for high sensitivity.
• Generating a high-quality, host-specific MS spectral library for accurate HCP identification.
• Data processing and label-free quantification of HCP peptides.
• Case Study: Implementation of SWATH-MS to monitor over 200 HCPs simultaneously during DSP scale-up.

Module 6: Targeted Quantification of Problematic HCPs (p-HCPs) using MRM

• Identifying and prioritizing p-HCPs based on biological risk.
• Methodology of Multiple Reaction Monitoring (MRM) and Parallel Reaction Monitoring (PRM).
• Optimizing peptide transitions, collision energy, and retention time windows for ultra-low LOD/LOQ.
• Developing a quantitative MS assay for GMP lot release
• Case Study: Designing an MRM assay to track a residual CHO cell protease known to cleave the final drug product.

Module 7: Bioinformatic Tools and Deep-Profiling Data Analysis

• Peptide identification, false discovery rate (FDR) calculation, and database searching.
• Software tools for HCP deep-profiling and data visualization
• Correlation of HCP data with purification steps to identify process bottlenecks.
• Statistical analysis of HCP variance for batch-to-batch consistency.
• Case Study: Using bioinformatics to determine the cellular location and function of the Top 10 most abundant residual HCPs.

Module 8: Integrating HCP Control into Quality by Design (QbD) and Risk Assessment

• Defining HCP control as a Critical Process Parameter (CPP).
• Developing a risk-based control strategy for HCP clearance throughout DSP.
• Tools for risk assessment.
• Establishing a design space for HCP reduction steps.
• Case Study: Applying QbD principles to justify a simplified DSP step based on HCP clearance data.

Module 9: HCP Clearance Study Design and Execution

• Designing spiking studies to evaluate HCP removal efficiency for different DSP unit operations.
• Mass balance and recovery calculations for accurate HCP monitoring across process steps.
• Scale-down model qualification and comparability to commercial scale.
• Regulatory expectations for HCP clearance data package in BLA submissions.
• Case Study: Demonstrating robust HCP removal by Protein A chromatography and polishing steps using MS data.

Module 10: Special Considerations for Gene Therapy Products

• Unique challenges of HCP analysis in Viral Vectors and cell-based therapies.
• Addressing the complexity of HCP encapsulation or co-purification with the vector capsid.
• Adaptation of LC-MS protocols for low-titer/high-complexity samples.
• Analytical approaches for monitoring residual plasmid DNA (rDNA) and other process impurities.
• Case Study: Developing a SWATH-MS workflow to profile HCPs in a GMP AAV drug product.

Module 11: Lifecycle Management and Process Changes

• Impact of upstream changes on the HCP profile.
• Assay lifecycle management.
• Strategies for HCP comparability studies following manufacturing site transfers.
• Handling lot-to-lot variability in both HCP levels and HCP profile.
• Case Study: Justifying the comparability of two drug substance lots following a cell culture media switch using both ELISA and MS.

Module 12: Automation and High-Throughput Screening

• Integrating automated liquid handling systems for HCP sample preparation and ELISA.
• High-throughput LC-MS method development for early DSP screening.
• Emerging biosensor-based assays and other rapid HCP detection methods.
• Data management and LIMS integration for regulatory audit trails.
• Case Study: Implementing an automated ELISA platform to increase QC throughput by 50%.

Module 13: Non-Immunological Orthogonal Methods

• Principles of Capillary Electrophoresis (CE) and HPLC for HCP separation and quantitation.
• 2D-DIGE and differential gel electrophoresis for qualitative profiling.
• Protein aggregation and HCP interaction studies using SEC-HPLC.
• Advanced sample preparation techniques for MS.
• Case Study: Utilizing 2D-DIGE to visualize the difference in HCP profiles between null and production cell lysates.

Module 14: Data Integrity and Regulatory Documentation

• GMP requirements for analytical methods and validation of LC-MS and ELISA.
• Ensuring data integrity for HCP results.
• Structuring the HCP control strategy section for IND/BLA filings.
• Responding to regulatory deficiencies related to HCP analysis.
• Case Study: Review of a successful BLA HCP data package focusing on MS supplementation.

Module 15: Future Trends in HCP Analysis and Personalized Medicine

• The role of Artificial Intelligence (AI) and Machine Learning (ML) in HCP data interpretation.
• Advancements in bioinformatics for immunogenicity prediction based on HCP sequence.
• Emergence of MS-only release assays for HCP quantification
• Integration of HCP analysis into high-resolution product characterization.
• Case Study: Exploring the use of AI to find correlations between DSP parameters and HCP clearance efficiency.

Training Methodology

This course employs a participatory and hands-on approach to ensure practical learning, including:

• Interactive lectures and presentations.
• Group discussions and brainstorming sessions.
• Hands-on exercises using real-world datasets.
• Role-playing and scenario-based simulations.
• Analysis of case studies to bridge theory and practice.
• Peer-to-peer learning and networking.
• Expert-led Q&A sessions.
• Continuous feedback and personalized guidance.

Register as a group from 3 participants for a Discount

Send us an email: info@datastatresearch.org or call +254724527104 

Certification

Upon successful completion of this training, participants will be issued with a globally- recognized certificate.

Tailor-Made Course

 We also offer tailor-made courses based on your needs.

Key Notes

a. The participant must be conversant with English.

b. Upon completion of training the participant will be issued with an Authorized Training Certificate

c. Course duration is flexible and the contents can be modified to fit any number of days.

d. The course fee includes facilitation training materials, 2 coffee breaks, buffet lunch and A Certificate upon successful completion of Training.

e. One-year post-training support Consultation and Coaching provided after the course.

f. Payment should be done at least a week before commence of the training, to DATASTAT CONSULTANCY LTD account, as indicated in the invoice so as to enable us prepare better for you.