Upstream & Downstream Processing | BioXcellence | Boehringer Ingelheim
Upstream and downstream processing at Boehringer Ingelheim BioXcellence™
Experts in every stage of biopharmaceutical processing
At Boehringer Ingelheim, we work with a variety of different host cell and strain types on a daily basis. These include cell culture and microbial organisms (bacteria and yeast), producing a wide range of molecule types such as:
- monoclonal antibodies,
- recombinant glycosylated and non-glycosylated proteins,
- peptides,
- antibody fragments,
- Fc fusion proteins,
- protein scaffolds and
- plasmid DNA (pDNA).
Our platform solutions for monoclonal antibodies and tailor-made solutions use a large toolbox of biopharma manufacturing techniques, but most of them fall into two basic categories: upstream and downstream processing.
The illustration below provides an overview of the biopharmaceutical production process based on mammalian cell culture. It maps each stage – from cell preparation to quality control – and shows how upstream processing transitions into downstream processing.
Example for cell culture biomanufacturing processes
What is upstream processing in biopharmaceutical manufacturing?
Upstream processing is the production of the active pharmaceutical ingredients (API) or active substance of biopharmaceuticals in a production bioreactor, the largest of which can hold a volume of up to 18,000 liters. In other words, the production of biopharmaceuticals typically starts with the ‘upstream process’– the generation of the API via living cells. It begins with mere milliliters of mammalian or microbial cells engineered to produce a specific molecule. Step by step, in the controlled environment of a bioreactor, these cultures are grown to a volume of several thousand liters to produce several kilograms of the desired API.
Differences in upstream processing of mammalian cell culture and microbial organisms
Mammalian cell culture is used to produce large, complex proteins like monoclonal antibodies. Using them allows for changes to the API – like those which may happen in human cells: glycosylation, for example. Mammalian cell production processes take several days or even weeks; growing them requires a complex and well-balanced mixture of nutrients as well as relatively gentle stirring and aeration. (Learn more about our [mammalian expertise](/content/partnering/boehringer-ingelheim-bioxcellence/mammalian-expertise "Explore our mammalian technology"/index.html).)
Microbial organism cultures can grow much faster, in days or even hours, producing smaller molecules like peptides, enzymes, antibody fragments and protein scaffolds very efficiently. Microbial organisms are usually very robust and can grow to high cell densities, generally, several orders of magnitude beyond mammalian cell cultures. However, they typically have some limitations on the size of the protein and 'human-like' modification of the molecules. (Read more about our [microbial expertise](/content/partnering/bioxcellence/our-business/microbial-expertise "Learn more about our microbial technology experience"/index.html).)
After the production of the desired molecule with the best suited organism via upstream processing, the product is then separated from a complex mixture of host cell proteins (bacteria and yeast), cells, cell debris, nutrients, and waste materials – which is known as the ‘downstream process’.
What is downstream processing in biopharmaceutical manufacturing?
The downstream process starts with separating the cells from the media. In most mammalian cell cultures, the protein is secreted into the media, and is collected for further purification. In microbial production, the protein can also accumulate inside cells, which need to be disrupted, and the cell debris separated from the API. Insoluble product agglomerations known as inclusion bodies – including unfolded, pure proteins sticking together – must be refolded into the biologically active conformation.
Various techniques are available to purify the API, but chromatographic separation is widely used in downstream processing. Separation principles used for chromatography are based on charge, size, hydrophobicity, or by specific affinity of the API to the chromatographic resin. Impurities can also be removed using precipitation based on temperature, pH, or chemical interaction. Moreover, a wide range of filtration techniques are used to separate the protein from the remaining solids, remove impurities, and exchange buffers.
Experts in upstream and downstream processing
At Boehringer Ingelheim BioXcellence™, we have decades of experience in developing, transferring and scaling up of biological processes, as well as in the utilization of cutting-edge modeling tools and state-of-the-art analytical methods. That's why we can offer efficient platforms and customized manufacturing solutions for a broad range of molecules, while maintaining secure supply of material throughout the whole lifecycle of an API. We're your trusted CMO/CDMO partner.