Validation of LC–MS Multi-Attribute Method Supporting Biopharma Process Characterization

The Microbial Process Development Group at Merck KGaA was tasked with developing and producing recombinant proteins expressed in Escherichia coli and Pichia pastoris. They used mass spectrometry (MS) as a routine tool for supporting process development in recombinant protein production. However, routine use of Multi-attribute method (MAM) in this environment meant overcoming various scientific, technological, and methodological challenges. These challenges included managing large amounts of data, producing unbiased audited results, and meeting process validation requirements. The overall production process for recombinant proteins involves multiple processing steps that are driven by defined process parameters. The characteristics of the resulting protein product must be such that the final drug substance is both safe and efficient. Studying the desired protein characteristics, then defining, monitoring, and managing critical quality attributes (CQAs) is key to success.

Merck KGaA is a major biopharmaceutical manufacturer based in Martillac, France. The company's Microbial Process Development Group is responsible for developing and producing recombinant proteins expressed in Escherichia coli and Pichia pastoris. The group includes scientists and experts in microbial processing analytics. They use mass spectrometry as a routine tool for supporting process development in recombinant protein production and are involved in developing original mass spectrometry-based approaches that enable better understanding of protein expression in microorganisms for biopharmaceutical development. Once a process is developed, it can be transferred to a manufacturing unit, where GMP specialists manage production, from pre-clinical stages through to commercial batch manufacturing of a drug substance.