Category Archives: Upstream Bioprocessing

Balancing Risk, Cost and Speed During Clinical Development While Still Maintaining Quality

In this podcast, we talked with Thierry Cournez Vice President, BioReliance® End-to-End Solutions, MilliporeSigma.. We discussed effective ways for emerging biotechs to collect material quickly and cost-effectively for pre-clinical and clinical studies. We also discussed managing the need to move quickly with cost and quality.

Show Notes

I began the interview by asking Mr. Cournez if he could talk about how MilliporeSigma works with emerging biotechnology companies. He said that developing and implementing a clinical material process can be time consuming and complex. He explained that MilliporeSigma is a CDMO partner for startups and small biotechs that are developing commercial biological drugs. They help these companies balance speed and cost by providing a comprehensive suite of products and services to accelerate clinical biopharmaceutical development, scale up production processes, and design and implement single-use commercial production facilities. In addition, they do not just focus on the technical and quality phase of drug development,  they also prepare supporting information for regulatory agencies. They make sure companies get all the support they need to explain to agencies what is in the dossier and answer their questions. Most importantly, they want to make sure that their customers are successful.

Next I asked Mr. Cournez how working with a CDMO is different than working with multiple suppliers. He said that after finalizing their strategy, small biotechs need to decide on how they want to structure their team. They have to ask whether they have all the expertise they need in-house or if they need to work with a partner to cover the considerable needs of process development, chemistry, manufacturing, control, filling and regulatory affairs. Many small biotech companies will choose to work with a partner and choosing a partner should not be taken lightly. Mr. Cournez said he feels it is key to the success of the project. Ideally the CDMO partner can interface with most of the individual suppliers and cover most of the requirements, but the biotech company does need an in-house person to work with the CDMO. At MilliporeSigma, they have a one dedicated project manager for each of their clients.

I then asked how small biotechs can collect material quickly and cost effectively to reach the pre-clinical stage. He said that this is where a CDMO can offer a real benefit. At MilliporeSigma, he explained, they learn from previous projects where time and money can be saved. He said they recently identified a two month time savings on the development of a molecule for a client.

Next we talked about how a CDMO can be instrumental during analytical development. He explained that analytical development is a critical part of the development process. Molecules can be really complex with no way of knowing what issues may arise. This is why analytical methods development and process development must work side by side to create a seamless process.

I then asked Thierry about what he thinks are the key considerations that small biotechs need to consider when working toward clinical development. He said that clinical development for a biotherapeutic is long and very challenging. Companies need to move quickly through the early clinical phases while demonstrating direct safety and efficacy. At the same time companies need a reliable process for producing clinical materials that ensures they will meet all quality and regulatory requirements. Long term, companies need to consider the final cost of their drug at commercial scale. Because of the complexity of this process, relying on an experienced partner can really help navigate these decisions early and be informed of everything that could impact decisions down the road.

I then asked how companies can balance cost, risk and speed during clinical development while still maintaining quality. He said that biotech companies all want the process done well, cheap and fast, but under no circumstances can we sacrifice on quality. MilliporeSigma employs a flexible approach to the biotech company’s needs and constraints. Again experience is very important as skilled partners can suggest new approaches and solutions. An example of an innovation through experience that has been launched is MilliporeSigma’s integrated plug and play upstream development services. This service eliminates the need to work with vendors for upstream development, thereby reducing bottlenecks and lowering time to clinic by 3 months.

I closed the interview by asking if there was anything else that Thierry would like to add for listeners and he said that these are very interesting times with many novel therapeutics in development. Many groups are working together to get these products into the hands of patients. At MilliporeSigma, we are looking forward to these collaborations.

Rapid At-line Media Analyzer Speeds Process Development by Eliminating Analytics Bottleneck

In this podcast, we talked with Dr. Glenn Harris,  Director of Integrated Life Sciences Platforms at 908 Devices, about the benefits and challenges of implementing rapid media analysis in process development, including the bottleneck created by outsourcing samples to core labs. We also discussed an easy to implement, benchtop media analyzer that permits comprehensive media analysis in real-time, thus speeding process development efforts.

I started the interview by asking Glenn if he could explain the benefits of rapid media analysis and also common pain points. He said that because spent media analysis usually can’t be done at-line, samples need to be sent out, and this creates a slow turn-around time. It also creates a logistics and tracking burden.

In addition, multiple technology platforms are required to run a sample, each having separated prep protocols and different sample volume requirements. This is particularly challenging with microbioreactor systems like ambrs or shake flasks, where labs contend with processing a large number of samples promptly coupled with sample volume constraints. The microbioreactor systems have small working volumes that limit the sample volume that can be removed each day to run samples. Since they permit several different media configurations to be run simultaneously, this results in a large number of samples at low sample volumes.

With at-line spent media analysis, you can run samples daily, or more often depending on your process. This provides rapid data day-to-day, hour-to-hour on the metabolic profile and media profile of the culture. Culture changes can be made on the fly, and processes can be optimized faster.

We discussed how in September, 908 Devices launched the Rebel® analyzer to address challenges in spent media analysis. Glenn shared what led the company to develop the Rebel and what key features were essential to include. He said that two years before building the Rebel, they began speaking with current customers in biopharma process characterization about their needs concerning spent media analysis. This meant getting in the lab with customers to see and experience the good and the bad of their day-to-day responsibilities. They identified two huge bottlenecks in process development, the number of samples that were being sent out, and the data coming back in (often in different propriety data formats). Both were extremely time-consuming to deal with.

908 Devices realized that the analyzer needed to be run alongside bioreactors in the process development lab. Current analyzers were often adapted from the clinical space, and users desired analyzers that were fit for purpose. From there, 908 Devices applied their super simple philosophy of design to the Rebel. This meant no computer on the side and no waste or solvents on the table. Putting everything inside the analyzer freed bench space and reduced footprint. Also, there were no special power requirements, no extensive sample prep, and the Rebel was 21 CFR part 11 compliant out of the box. With its small footprint, the Rebel fit under shelves and was easily mobile on carts.

Next, I asked how the Rebel could help speed process development efforts. Glenn said that they focused on making sure that the Rebel could be positioned where samples were originating to ensure at-line analysis. This eliminated the need to send samples out of the lab. Also, sample prep had to be super simple, with no special prep or labeling required. Once samples are put into the system with either vials or well plates, the system provided unambiguous data on what was detected and the concentration level. The data was not in a proprietary format, instead provided as a CSV or pdf file. This data could be delivered on the device or a mapped network drive.

I asked how users would implement the Rebel in their labs and if there was any specific training needed. Glenn explained that the Rebel sits alongside bioreactors, so when a sample is pulled, you just send a little bit to the Rebel. The training is short, typically a half-day or less. He shared that they have trained 2 – 20 researchers in a day on everything needed to use the Rebel with approximately 90% in hands-on time training. They also provide a dedicated support team with engineers, scientists, and data science gurus to answer any follow-up questions once they leave.

We then discussed the user experience. Glenn said that everyone who could reach into his or her pocket and unlock his or her phone could run the Rebel. It has a large touch screen interface, simple notifications, limited external interfaces, no external keyboard, and minimal maintenance. He said that they worked hard to make interactions with the Rebel a breeze since science in the process development lab is hard enough.

I asked if the Rebel could be used in cell and gene therapy applications. He said that cell and gene therapy groups are utilizing the Rebel to optimize media for their applications to ensure reproducible processes, increase yield, and improve media and product quality. Many of the cell and gene therapy companies said that they are learning from the biologics community about how important media is and as a result, are working closely with media providers to ensure consistency

Next, I asked about the feedback from customers. Glenn said that fortunately, they have seen overwhelmingly positive responses thus far. He said it was great to see systems being used as intended soon after they left installations and trainings. It was good to see that users were confident using the system. They are also seeing many orders coming in for more consumables as teams realize the benefits of media screening and optimization in near-real-time. Users have been responsive to emails and check-in calls and love to share internal success stories with their colleagues.

I asked if there have been any challenges associated with the implementation of the Rebel. Glenn shared two that came to mind. The first was networking. He explained that there has been a big push for Pharma 4.0, yet some IT departments are resisting network enabling of devices in labs. Luckily, researchers and engineers have been a great ally and have been in full support of implementation.

Another challenge has been the output of data. Researchers that were used to waiting days or weeks for data are now getting it in minutes. He said that “it surprised some groups when they loaded up a tray of samples and came into the lab the next day with nearly 100 files of time course spent media data from their processes”. “We are working with these groups on streamlined, no-fuss data analysis strategies to keep up with their ever-growing sample sets and curiosities.”

I closed by asking if Glenn had anything else to add for listeners. He said that collaborations are critical, both public and private. One in particular with Sarah Harcum’s group at Clemson University has been spectacular since they are key opinion leaders in the process development space. They just published their first collaborative work looking at spent media analysis in ammonia stressed CHO cultures grown in an ambr 250 microbioreactor platform.

908 Devices is also looking toward future development, new apps, and platform expansion with collaborators on a NIIMBL project. He added that listeners should feel free to reach out to him directly on email through or LinkedIn for any additional information.

Successful Methods for Perfusion Process Optimization

In this podcast, we interviewed Dr. Andreas Castan, Principal Scientist at GE Healthcare Life Sciences about the best methods for optimizing perfusion processes. This included a discussion of tools for media optimization and innovative cell separation techniques.

Show Notes

I began the podcast by asking Dr. Castan what makes perfusion a good manufacturing platform? He identified several reasons including short residence times in the bioreactor through adding fresh media and removal of spent media and as a result is perfect for unstable molecules. Perfusion permits processes to be run with high volumetric productivity by maintaining high cell densities for a long period. With perfusion there are many opportunities for process intensification and it is a good way to quickly get started with production, provided you have a good batch media that cells like.

Next I asked if there were any specific product types or situations that are an especially good fit for perfusion? Andreas described three primary areas – unstable products, integrated continuous manufacturing platforms and process intensifications for removing bottlenecks in manufacturing. Examples would be cell bank manufacturing to create high density cell banks, seed train to remove steps or produce high viable cell density inoculum or hybrid processes of both perfusion and fed batch technologies.

We then discussed the importance of optimization for perfusion processes and the various approaches for optimization. Andreas said it is important to optimize processes to gain the product quality and the process economy that you are aiming for. Cell culture media is the most important thing to optimize in perfusion culture. In order to achieve good throughput for your optimization it needs to be performed in a scale down model then results must be verified under bioreactor conditions.

Then I talked to Andreas about a recent poster he authored and presented at ESACT. The poster outlines development of perfusion specific media and designing medium that supports low cell-specific perfusion rates. I asked him to explain what cell-specific perfusion rate (cspr) is and why a low cell-specific perfusion rate is important? Andreas explained that cell-specific perfusion rate is the volume of media added per cell per day. If you assume your medium supports a cspr of 50 picoliter and you want to run the process at 30 million cells/mL your volumetric perfusion rate would be 1.5 bioreactor volumes per day. If you take the same medium with 100 million cells/mL you would need a volumetric perfusion rate of 5 bioreactor volumes per day, which is not feasible for production. A medium with a better depth of cspr of 10 picoliter at a cell density of 100 million cells/mL would have a volumetric perfusion rate of 1 bioreactor volume per day, which is feasible for production.

His team investigated screening methods to develop medium with a very low cspr. They started with basal media, then screened different feed solutions and Cell Boost feeds in batch mode. The Cell Boosts with a positive impact were taken to a DOE study in spin tubes that replicated perfusion conditions, then promising formulations were moved to perfusion processes in WAVE or the XDR bioreactors.

Poster – Perfusion media development for scalable processes

Next I asked him if you can turn a fed-batch medium into a perfusion medium? He said yes, transferring a fed batch media to a perfusion process is a simple process. He lays out the methodology in the previously mentioned poster and they have done this for two cell lines and three fed-batch media and feeds. The methodology is also described in the Biotechnology Process Journal Article, “Repurposing fed‐batch media and feeds for highly productive CHO perfusion processes.”

I followed up by asking how did the methodology work in the case study presented in the poster? Andreas said it was a very fast method with batch screening taking one week, the DOE study took two weeks and within one month they had the composition for a perfusion media that fit the desired clone. They tested two media developed using this process on their internal Herceptin producing cell line and for both media they reached cspr below 20 picoliter. They were able to reduce one media to only 7 picoliter. The cell specific productivity for this process was comparable to what they saw in fed-batch.

We then discussed another approach to optimizing perfusion processes, optimizing the equipment used in process. In another poster presented at ESACT, Andreas presented the use of hydrocyclones for cell separation. I asked him to describe how hydrocyclones work and why they make an attractive alternative to traditional cell separation devices? He said that hydrocyclone devices are comprised of cylindrical and conical parts that allow centrifugal separation provided by feed suspension introduced tangentially at high flow rates into the device. The absence of rotors or moving parts make it an interesting separation alternative to perfusion in long term operation. A simple device that is not prone to clogging as is usually seen with filtration devices

Poster – Hydrocyclone for mAb production in a perfusion single-use bioreactor

I then asked what his experiences were with the hydocyclone in the case study? Andreas explained that they investigated separation efficiencies at different flow rates and different flow concentrations and found that separation efficiencies of 70-80% could be achieved with the current hydrocyclone. This test was run using a perfusion process in an XDR 50 bioreactor at 50 million cells/mL for more than 2 weeks in very stable conditions. It was a very successful test of the device.

I summarized by asking if companies are limited in time and/or resources for optimizing their perfusion processes, what would he focus on first? He said media should be focused on first because as explained earlier a low cspr is key to reaching high volumetric productivity, low volumetric perfusion rates and high product concentrations, all of which result in good process economies. Next you could think about how you could use perfusion to reduce process scale.

Lastly, I asked if he had anything else that he would like to add for our listeners. He said that before moving into perfusion you should ask yourself what makes the most sense for your process. You need to think about your infrastructure, prerequisites, platform, and previous knowledge. Then you can decide where perfusion makes the most sense in your process. In cell bank manufacturing, seed train, or production bioreactor. Media is the most important factor to work with and it is not difficult to use existing media and cell boosts to develop perfusion media that fits your clone.