Monitoring process-related impurities in biologics–host cell protein analysis

Abstract

During biologics development, manufacturers must demonstrate clearance of host cell impurities and contaminants to ensure drug purity, manufacturing process consistency, and patient safety. Host cell proteins (HCPs) are a major class of process-related impurities and require monitoring and documentation of their presence through development and manufacturing. Even in residual amounts, they are known to affect product quality and efficacy as well as patient safety. HCP analysis using enzyme-linked immunosorbent assay (HCP-ELISA) is the standard technique, due to its simple handling, short analysis time, and high sensitivity for protein impurities. Liquid chromatography mass spectrometry (LC–MS) is an orthogonal method for HCP analysis and is increasingly included in regulatory documentation. LC–MS offers advantages where HCP-ELISA has drawbacks, e.g., the ability to identify and quantify individual HCPs. This article summarizes the available knowledge about monitoring HCPs in biologics and presents the newest trends in HCP analysis with current state-of-the-art HCP measurement tools. Through case studies, we present examples of HCP control strategies that have been used in regulatory license applications, using an MS-based coverage analysis and HCP-ELISA and LC–MS for HCP quantification. This provides novel insight into the rapid evolving strategy of HCP analysis. Improvements in technologies to evaluate HCP-ELISA suitability and the implementation of orthogonal LC–MS methods for HCP analysis are important to rationally manipulate, engineer, and select suitable cell lines and downstream processing steps to limit problematic HCPs.

What is the role of current mass spectrometry in pharmaceutical analysis?

“…A monitoring process-related impurity in biopharmaceuticals was extensively discussed in a recent review (Pilely et al, 2022). The advantages of LC-MS techniques over enzyme-linked immunosorbent assay (ELISA), for example, the ability to identify and quantify individual host cell proteins, were summarized through case studies.…”

Saccharomyces Specific- HCP Assay for r-PAI-2 Therapeutic Proteins.

“…There are new approaches that are applicable for understanding residual HCP that can facilitate assessments of safety risk and potentially lead to better control of process impurities in the therapeutic products. Recent developments included liquid chromatography mass spectrometry (LC-MS) which are current state-of-the art tools for HCP measurement [23,25].…”

Establishment and evaluation of detection methods for process‐specific residual host cell protein and residual host cell DNA in biological preparation

“…A method for detecting with high sensitivity and good repeatability is not only the key to ensure the safety and effectiveness of biological products, but also an important parameter for the control of the production process and process optimization. Nowadays, the main detection methods for HCP include enzyme‐linked immunosorbent assay (ELISA), SDS‐PAGE silver staining, HPLC, capillary electrophoresis, and mass spectrometry, 15–19 all of which have certain advantages and disadvantages. ELISA has been widely used due to its good specificity, high sensitivity, quantitative ability, and objective judgment of the endpoint.…”

“…Nowadays, the main detection methods for HCP include enzymelinked immunosorbent assay (ELISA), SDS-PAGE silver staining, HPLC, capillary electrophoresis, and mass spectrometry, [15][16][17][18][19] all of which have certain advantages and disadvantages. ELISA has been widely used due to its good specificity, high sensitivity, quantitative ability, and objective judgment of the endpoint.…”