Large-Scale Assessment of Extractables and Leachables in Single-Use Bags for Biomanufacturing

Area of Science:

• Biopharmaceutical Manufacturing
• Analytical Chemistry
• Materials Science

Background:

• Single-use technologies (SUTs) are integral to modern biopharmaceutical manufacturing, offering advantages in flexibility and reduced contamination risk.
• However, the potential release of extractable and leachable (E&L) substances from SUTs, such as single-use bags (SUBs), poses a significant concern for drug product safety and efficacy.
• E&Ls can impact cell culture performance, complicate purification processes, and potentially harm patients if present in the final drug product.

Purpose of the Study:

• To comprehensively analyze the extractable and leachable profiles of 34 plastic films from SUBs used in cell cultivation.
• To investigate the influence of bag features (brand, manufacturing date, polymer type) and manufacturing changes on E&L profiles.
• To generate a robust E&L database to aid in toxicological assessments and guide the selection of suitable materials for bioprocessing.

Main Methods:

• Plastic films from 34 SUBs were extracted under simulated worst-case biomanufacturing conditions using accelerated aging.
• Analyte extraction and preconcentration were performed using dispersive liquid-liquid microextraction (DLLME).
• Extracts were characterized using GC-headspace (volatiles), high-resolution GC-Orbitrap-MS/MS (semi-volatiles), high-resolution LC-Orbitrap-MS/MS (non-volatiles), and ICP-MS (trace elements).
• Multivariate statistical analysis was employed to correlate E&L compounds with bag characteristics.

Main Results:

• Significant correlations were identified between the types and concentrations of E&L compounds and specific bag features, including brand, manufacturing date, and polymer composition.
• The study revealed that the E&L profiles of SUBs have evolved over time due to manufacturing modifications and the adoption of new polymer types.
• A diverse range of volatile, semi-volatile, non-volatile, and elemental E&Ls were detected and characterized.

Conclusions:

• The E&L landscape in single-use bioprocessing is dynamic, influenced by manufacturing practices and material innovations.
• The generated E&L database provides critical data for toxicological risk assessments in biopharmaceutical manufacturing.
• These findings offer practical guidance for screening and selecting appropriate SUB films, ensuring the safety and quality of biopharmaceutical products.