You Need to Reach Preclinical Phase? What Now?

The prime goal of an early-stage biotech company after securing funding and implementing the first steps – deciding on a strategy and understanding risks – is to collect material as quickly and cost-effectively as possible in order to reach the preclinical phase. However, many companies can be so blinded by this goal that they may overlook potential stumbling blocks along the way, a mistake that can sometimes cost a company its existence.

“Getting it right the first time is essential,” says Michael Lanero Fidalgo, Director, Head of EMEA Biodevelopment Center at Merck KGaA*, Darmstadt, Germany. “There is no second chance. You need to get through the preclinical phase to reach phase I. And your phase I clinical trial is your reference for the future and validates the proof of concept of your proposed new therapy.”

The upstream phase is absolutely critical for long-term success. This, in turn, emphasizes the importance of getting it right the first time. “Many early-stage companies think they can easily redo things if they go wrong, but what they do not realize is that this will cost them a lot of time and money,” Lanero Fidalgo explains.  

The key, he says, is to control quality. “It is essential to find the best compromise at an early stage to reduce risk and keep the costs down at the same time. Sometimes it is necessary to invest in quality and effort at the beginning, so as to speed up the process in the end.”

Finding a partner to reach the preclinical phase

Lanero Fidalgo, Merck, preclinical phase, Bioreliance End-to-End Solutions

Michael Lanero Fidalgo, Director, Head of EMEA Biodevelopment Center at Merck KGaA*, Darmstadt, Germany

Overall, gaining a foothold in the industry and preparing the process to develop a promising product can be quite overwhelming for early-stage or small biotechs. Here, a partner can be of help. At early stages, many biotechs turn to a trusted CRO or CDMO to guide them through the different stages of drug development and to bring the necessary expertise and infrastructure to the table.

“When choosing a partner, biotechs need to consider two things,” says Lanero Fidalgo. “First, a partner needs to be an extension of the team, they need to be trustworthy and adaptable in case of issues. Second, they need to prove that they have expertise in the field the biotech company is working in. When looking for a partner, the biotech should therefore consider the successful track record of CROs or CDMOs.”

Lanero Fidalgo is part of Merck KGaA*, Darmstadt, Germany, and focuses on BioReliance® End-to-End Solutions. The portfolio of services from BioReliance® End-to-End Solutions supports companies during all stages of development. Their Plug & Play Upstream Development Service is tailored to small biotech companies that need to balance cost, risk, and speed on their way to the clinic while getting it right the first time. The adaptive approach is made up of services modeled to the needs of small and emerging biotechs from DNA, through the preclinical phase, to the clinic.

“Biotechs usually all want the same: they want the process to be done good, cheap and fast,” he explains. “But under no circumstances do we compromise on quality. We always try to be flexible and tailor our approach to the biotech’s needs and constraints. Our experience here is important because we can propose feasible approaches and often shortcuts that have already worked on other projects.”

So what about the science? The mini-pool approach

mini-pool approach, Merck, laboratory work, cell line development, clone selection

By learning from past projects, the team that offers BioReliance® End-to-End Solutions is constantly adapting, tailoring and optimizing its approach to drug development, and understanding where money and time can be saved. “Each project is different,” says Lanero Fidalgo. “But there are similar strategies that work for different projects. For example, depending on the risk a biotech is willing to take, we offer our  recently developed, automated mini-pool approach as a fast-track process, which can save up to two months.”

This approach allows the team to save time through the selection of mini-pools displaying the best titers and growth profiles at the early cell line development stage. The selected pools are used as starting material for process development, while the clone selection continues in parallel. The best conditions are confirmed using the final clone.

“We use the mini-pool approach for cell line development,” adds Cyrielle Corbin, Team Leader, European Biodevelopment Center at Merck KGaA, Darmstadt, Germany. “Here, the diversity is significantly reduced, giving the greatest chance to the best performing clones. It is expected that clones emerging from the cloning step of the mini-pool will display similar cell behavior in the bioreactor as in the mini-pool.”

Cell line development with CHOZN® GS

Cyrielle Corbin, Merck, preclinical phase, Bioreliance End-to-End Solutions

Cyrielle Corbin, Team Leader, European Biodevelopment Center at Merck KGaA, Darmstadt, Germany

To increase the chances of selecting the right clone, the team supporting BioReliance® End-to-End Solutions uses the so-called CHOZN® GS platform, which is based on the commonly used and stable CHO (Chinese Hamster Ovary) cell line and provides stable cell lines that produce the protein of interest.

“The CHOZN® platform is characterized by a depletion of the endogenous genes encoding the glutamine synthetase enzyme,” explains Corbin. “When we introduce the gene of interest as well as the glutamine synthetase gene into the cells, we do not need to add glutamine to the culture medium anymore. So by simply removing the glutamine from the culture medium, we are able to select the cells that have actively integrated the glutamine synthetase genes. And this usually correlates with the integration of the gene of interest.”

In short, the removal of glutamine synthetase provides a selective system that facilitates the elimination of those cells that did not take up the glutamine synthetase gene and the gene of interest, and therefore do not have the ability to survive without glutamine. This depletion process requires no added chemicals and will result in highly stable and productive clones for the preclinical phase.

Selecting the right clone

mini-pool approach, Merck, laboratory work, cell line development, clone selection, preclinical phase

“When it comes to selecting the right clones, we typically have 20 clones at the final screening phase,” Corbin explains. “We monitor their cell growth, metabolism and productivity performance for 14 days, and if quality specifications have already been defined by the customer, we also analyze the recombinant protein produced by each of these clones.”

The best clone, says Corbin, should fulfill three specific criteria. First, if quality specifications are already defined, the quality of produced protein should be as expected. Second, the clone should have a high productivity performance, for example, a high titer or high cell-specific productivity (Qp). Lastly, the best clone should display the best viable cell density (VCD) and good viability at the end of the productivity assessment.

“The viability is very important for process development because we have to ensure that the process development will face the least trouble moving forward,” Corbin says. “If the customer wants to increase the titer, for example, good viability will allow us to pursue the productivity assessment for several days, which will result in an increase of the titer.”

If not all of the three criteria can be found in a single clone, a compromise between the three criteria will help the researchers determine the best clone amongst the 20 available clones. Most importantly, the clone has to be stable in terms of performance and quality during the entire production phase. Clone stability is confirmed by a dedicated study that is performed on several clones. Combining all of these results, the team with BioReliance® End-to-End Solutions supports the customer in selecting the right clone for the production of their protein.

Combining multiple workstreams for efficiency

mini-pool approach, Merck, laboratory work, cell line development, clone selection

BioReliance® End-to-End Solutions has set up a process in which multiple workstreams within a project can run in parallel, including cell line development, clone selection and stability, pre-material production, and analytical development.

“If the customer is really in a hurry, we might also propose to start on process development, while also working on the clone selection phase of cell line development,” Corbin explains. “Moreover, during the clone stability phase, we can also proceed to master cell bank (MCB) manufacturing. As we are using the CHOZN® GS platform, we have a great chance of selecting the right clone. By proceeding in parallel we can save a lot of time.”

The analytical team and the cell line development team supporting BioReliance® End-to-End Solutions work side by side to ensure a smooth process to the preclinical phase. Within the team, a dedicated project manager and a technical leader drive the customer’s project, and one scientist is responsible for coordinating the activities during cell line development and keeping the analytical team and process development team informed.

This stringent collaboration ensures that the entire project is a success and that the early-stage biotech gets it right the first time. After all, there is rarely a second chance.

Interested in learning more? Check out the Plug & Play Upstream Development Service tailored to small biotech companies. If you are in the U.S. or Canada, click here! If you are in the rest of the world, click here!

If you are in the U.S. or Canada and want to learn more about BioReliance® End-to-End Solutions click here!

If you are in the rest of the world and want to learn more about BioReliance® End-to-End Solutions click here!

*The life science business of Merck KGaA, Darmstadt, Germany operates as MilliporeSigma in the U.S. and Canada.


Images via joker1991, Matej Kastelic, Viacheslav Lopatin/Shutterstock.com


Author: Larissa Warneck, Science Journalist at Labiotech.eu

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