The evolution of nucleotide production: a Roche CustomBiotech journey

mRNA vaccines and therapeutics harbor the potential to treat a broad range of diseases

Tapping into that potential hinges on efficient production processes that generate reliable and high quality products with minimized environmental impact.

At Roche CustomBiotech, we believe that sustainability and safety starts with the smallest molecules. For decades, we’ve pioneered novel approaches to designing and making a broad range of nucleotides (NTPs) that meet the diverse needs of biomanufacturing. That experience converges in our most recent innovation: exceptionally pure, animal-origin-free NTPs that conform to stringent quality standards and are produced without chemical solvents that pose environmental and safety risks.

From sledgehammer to scalpel: rethinking chemical production

In the 1990s, NTPs were typically produced in a chemical synthesis involving large quantities of organic solvents like DMF. DMF is not only difficult to replace but also prone to forming nitrosamines during chemical reactions.1 The presence of nitrosamines in pharmaceutical products is a documented global safety concern.2 These impurities are strictly regulated by authorities because they can evolve at any point in the synthesis of a drug and even trace amounts are genotoxic and mutagenic.3 Pyrophosphate was also a key reagent in the synthetic reaction, a known inhibitor of the polymerase chain reaction (PCR), and notoriously difficult to remove from the product.  


Dr. Hans-Georg Ihlenfeldt, Associate Director Operations at Penzberg, was among a group of scientists at Roche who saw the need for change already 30 years ago. He explains, “It was like a sledgehammer method, and what was needed was a process that worked more like a scalpel.” With long-standing expertise in enzymatics, Roche had the know-how and resources to make a difference. “We came up with a biological method using enzymes,” he says. “Why enzymes? Because they have been optimized over millions of years by nature to produce clean nucleotides efficiently. Enzymes were our scalpel.”

Our development and use of enzymatic processes removed organic solvents from manufacturing. “Sustainability in production is a driver of today’s innovations, but 30 years ago, as we established our biological method, we were far ahead of everyone in making our process green and environmentally sound,” states Ihlenfeldt.


Roche CustomBiotech is renowned for delivering high-quality reagents that meet stringent industry requirements. 

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"Sustainability in production is a driver of today’s innovations, but 30 years ago, as we established our biological method, we were far ahead of everyone in making our process green and environmentally sound".

Dr. Hans-Georg Ihlenfeldt
Associate Director Operations Penzberg & Former Developer

Modern processes and facilities for a safe and green supply chain

Ihlenfeldt and colleagues showed initiative and ingenuity in anticipating the need for improved NTP manufacturing. Today, that same spirit of innovation sparked another generation of innovations that further align our NTP offering with the standards of biopharmaceutical manufacturing. 

Collaborating with a multidisciplinary team of experts, Dr. Toni Pfaffeneder, Lab Head of Nucleotide Chemistry R&D, worked on those developments. He describes their motivation: “The enzymatic method for NTP production was a game-changer. The presence of pyrophosphate in the product was, on the one hand, a problem for PCR. On the other hand, it was akin to having a roadblock in the middle of a manufacturing highway. Downstream removal steps hampered the smooth process flow and could lead to suboptimal results. With enzymes, the roadblock was gone, and obviating organic solvents was also visionary. However, in today’s world, raw materials for the sensitive production of therapeutics must also be free from animal components.” 

NTP developemtent Roche expertise mRNA reagents

"It made sense to develop an advanced biotransformation process that is animal-origin-free with minimal risk for trace contaminants like nitrosamines. So, we got to work."

Dr. Toni Pfaffeneder
Lab Head Nucleotide Chemistry R&D

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aof antibiotics-free

The result of these efforts is an animal-origin-free (AOF) and antibiotic-free production process that eliminates concerns of nitrosamine contamination. Coupled with a brand-new manufacturing facility with customized automation technology designed exclusively for mRNA raw material requirements, Roche delivers consistently, high-quality NTPs with ensured safety and at scale.

Always striving to go the extra mile, the team also fine-tuned the process to systematically deplete impurities with high efficiency. That dedication underscores our commitment to a secure and green supply chain for biomanufacturing. “It is something that rarely stands in the foreground but is absolutely essential,” says Pfaffeneder. “Roche CustomBiotech has a longstanding and leading presence in the industry. Not only can you always count on the quality we provide today, but you can also expect that experts across all our departments are continually planning, developing, and improving products to stay at the forefront of technological advances and ensure that your production remains state-of-the-art.”

Well prepared to be part of a sustainable future

While enzymatic production of NTPs has been around for some time, the applications that use NTPs progress and change. It stands to reason that the features and manufacture of NTPs must follow suit. 

Today, we power NTP manufacturing with nature's own catalysts in a highly precise, environmentally friendly, and efficient process. Merging green chemistry with stringent quality standards, Roche CustomBiotech NTPs are fit-for-purpose and therefore tailor-made for safe biopharmaceutical manufacturing.

What about tomorrow? Untold technological advances are likely to transform how and what we create to ensure the world benefits from sustainable and safe biomanufacturing. Roche CustomBiotech will lead that transformation. In the words of Pfaffeneder: “Green chemistry and exceptional quality are key factors that set us apart and ensure that both we and our customers are well-prepared for the future.”

  1. European Medicines Agency, Committee for Medical Products for Human Use. 2020. Assessment Report: Nitrosamine impurities in human medicinal products. (Accessed May 2024)
  2. Vikram, H.P.R. et al. 2023. Nitrosamines crisis in pharmaceuticals – insights on toxicological implications, root causes and risk assessment: a systematic review. Journal of Pharmaceutical Analysis (in press). doi: 10.1016/jpha.2023.12.009
  3. Tuesuwan, B. and Vongsutilers, V. 2021. Nitrosamine contamination in pharmaceuticals: threat, impact, and control. Journal of Pharmaceutical Sciences 110: 3118. doi: 10.1016/j.xphs.2021.04.021