Monitoring specific process-related impurities

Regulatory authorities demand documentation for the efficiency of downstream purification of specific enzymes and proteins added during process development to ensure patient safety, drug stability, and efficacy.

Mass spectrometry analysis can detect all types of process-related protein residuals, e.g., affinity capture chromatography proteins, growth factors, Benzonase® nuclease, Protein A, enzymes for site-specific PEGylation, aminopeptidase, serum albumin, cytokines, protein-based media components, carrier proteins, and more.

Using LC-MS analysis with Multiple Reaction Monitoring (MRM), unique signature peptides for specific proteins can be identified and quantified even in small amounts. The assay is highly reproducible and, therefore, ideal for batch-to-batch comparison.

The LC-MS assay can be fully validated according to ICH guidelines and performed under GMP conditions as a release assay if needed.

ELISA (enzyme-linked immunosorbent assay) has been the gold standard in the industry for HCP analysis for many years. However, it is restrained by a lack of specificity and a limited possibility to monitor multiple specific proteins in highly complex samples. Developing a process-specific ELISA is time-consuming and typically takes 1 to 1.5 years.

A customized mass spectrometry (LC-MS) assay for MRM analysis can be developed and validated within six months. It may quantify even low amounts of specific proteins utilized during manufacturing.

We can measure up to 20 individual proteins in the same LC-MS run using multiplex analysis. The technique is highly reproducible and provides both the identity and absolute amounts of specific analytes. Furthermore, the method can be validated and qualified according to your specifications.

Polysorbate surfactants are crucial to the shelf-life and efficacy of monoclonal antibody (mAb) products, and their degradants may form particles that are harmful to patients and considered a critical quality attribute outlined by the USP General Chapters <787> and <788>. Consequently, degradation of polysorbates is a major problem in mAb development, resulting in project delays, often in very late stages of project development.

While some host cell proteins (HCPs) – such as proteases that cleave the therapeutic protein – are usually found early on, polysorbate-degrading lipases and esterases can remain undetected in the final drug product until months or years into long-term stability studies – or even after the product has been licensed!

At this stage, modifying the manufacturing process to eliminate these elusive yet dangerous HCPs is costly and time-consuming. Identifying polysorbate-degrading HCPs in a mAb product before they become a problem is the most effective way to reduce their impact on the project timeline.

Learn more in our Q&A PDF >>