Monthly Archives: May 2020

Bi-Specific Antibodies – The development, manufacture and promise of these cutting-edge therapeutics



In this podcast, we conducted a panel discussion with experts from Selexis and KBI Biopharma on bi-specific antibodies. We examined bi-specific antibody development and manufacturing, including current challenges and key solutions. We also discussed the promise of these cutting-edge therapeutics and their future in medicine.


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 www.908devices.com or LinkedIn for any additional information.


Non-Animal Origin cell culture supplements and manufacturing aids for biologics manufacturing



In this podcast, we talked with Dr. Tobias Hertzig, Regulatory Affairs Manager and Dr. Ulrich Tillmann, Global Product Manager, Supplements and Manufacturing Aids both representing Merck KGaA, Darmstadt, Germany. We discussed non-animal origin cell culture media supplements, including regulatory advantages, performance metrics and their use in biologics manufacturing.

I began our discussion by asking Tobias if he could give us a definition of what non-animal origin means and if there was an industry-wide understanding of the term. He explained that he would love to give an industry-wide definition for non-animal origin, but unfortunately there is no industry-wide definition and there is no useful or clear definition provided by the regulators.

Next I asked Tobias if he could talk about the distinctions between primary, secondary and tertiary levels for non-animal origin. He said that these terms are used to distinguish how far away from animal origin the material is. For example, primary describes something that is not directly derived from an animal source. Secondary describes, if you use fermentation as an example, that no animal sourced media ingredients were used in the manufacture. A good example for tertiary would be a recombinant insulin where the material is recombinant, the enzyme used to cleave the protein isn’t of animal origin and the media used to make the enzyme didn’t contain any product of animal origin. It is important to note that these terms aren’t fully defined so end users need to be sure that they understand what the author means by those terms as well.

I then asked Tobias to explain why animal origin is such a big concern in the cell culture realm. He said that the primary concern is that adventitious agents could be present in animal-derived material. However, other concerns could be based on a variety of issues, including: religion, kosher/halal, or lifestyle concerns, like allergies. For adventitious agents, there is concern about zoonotic agents that could cross from animals to humans and cause disease. There are several agents known to cross species, including prions such as TSE and BSE, and also viruses. The cells themselves can be susceptible to viruses and if any viruses are detected, then the manufacturing is not GMP compliant. There are also other concerns about animal-derived materials related to the supply situation including possible import restrictions and shortages.

I asked if he could share the regulatory view on non-animal origin components. He said it is preferred to avoid animal material whenever possible, but due diligence is always required even when non-animal origin is declared. It is important to understand why the material is considered non-animal origin and to work with suppliers to ensure that your understanding of non-animal origin is the same as their definition.

We then switched gears and I asked Ulrich if he could tell listeners about which products in the SAFC® portfolio are of non-animal origin. He explained that the non-animal origin supplements in their portfolio are recombinant proteins that mirror serum sourced versions. For example, instead of Fetal Bovine Serum (FBS), you have recombinant transferrin or albumin and for growth factors you have recombinant insulin or Long® R3. There are also manufacturing aids that are now recombinant, like trypsin for example. The portfolio of recombinant supplements and manufacturing aids is offered under the CellPrime® brand, so when you see that brand you know that it is a recombinant, non-animal origin supplement or manufacturing aid.

For clarification, I asked Ulrich to explain what Long® R3 is. He said Long R3 is a derivative of the IGF-1 growth factor that aids in growth of cells and prevents apoptosis. It also helps with utilization of nutrients in the media. It is similar in action to insulin, but receptors on the cells and their genetic make-up dictate whether they respond better to insulin or IGF-1. The ‘long’ in Long R3 IGF refers to a 13 amino acid N-terminal extension which aids in folding the peptide in E.coli. Glu3 (E) in the human IGF sequence has been replaced by Arg (R). The E to R substitution reduces binding to IGF sequestering proteins thus making the growth factor more available in the media.

I then asked how these supplements are used. Ulrich said that the supplements are being used as raw materials for manufacturing cell culture media. Recombinant insulin or Long R3 are either included in the media during manufacturing or are added by the end user. Media including these supplements are widely used for antibody manufacturing, in cell and gene therapy, and in vaccine and viral therapy media.

I then asked him to talk about mAb (monoclonal antibody) and recombinant protein manufacturing versus regenerative medicine, gene therapy and vaccine manufacturing with respect to supplement use. Ulrich explained that in mAb and recombinant protein manufacturing, companies are trying to reduce extraneous protein load in the media that complicate the purification of the desired product. So in those applications, only low molecular mass recombinant growth factors are used. In cell therapy, you still have many companies using serum components due to a lack of suitable replacements. In cell therapy, you still have many companies using serum components due to a lack of suitable replacements and people are only slowly getting around to using recombinant supplements in manufacturing instead. For instance, if you think about the extension of mesenchymal stem cells or adherent vaccine production cell lines, they are grown on supports from which the cells need to be split. In order to keep risk low, recombinant trypsin should be used instead of bovine or porcine based trypsin.

Next Ulrich described how just because something says ‘plant-derived’ on the label, it doesn’t necessarily mean that it is non-animal origin. It is important to closely look at their manufacturing processes. Where the plants are grown and under what kind of controlled conditions. If it isn’t tightly controlled, contamination can happen such as by animal manure.

We then talked about if there is a difference in performance between non-animal origin and animal origin supplements. Ulrich said that he doesn’t think that there is a difference in performance but with serum you have other issues, such as possible TSE/BSE contamination, introduction of proteins that you don’t want in your process, particularly as you move to purify the desired product. With differences between cell lines, it is important to titrate NAO supplements into cell cuture media in order to achieve the right concentration. MS supplies supplement sample pack sizes to those who make their own media and also offers custom media formulations (cell culture media containing supplements) to help companies get the best performance from their cell line.

I then asked Ulrich about the challenges in developing non-animal origin supplements. He said that you must be sure that you  have complete visibility on the manufacturing process. When you rely on suppliers for supplements and manufacturing aids, you must audit the suppliers and all the details of the materials they use and their manufacturing facility. You must assess their level of non-animal origin production. Products within the SAFC® portfolio use the definition that there can be no animal components in the raw materials used to formulate the supplement/manufacturing aids growth media; no animal origin material in their manufacturing process or in the facility; no shared equipment and no contact with animal origin used elsewhere in the manufacturing plant.

I closed our talk by asking if either Tobias or Ulrich had anything to add for listeners. Tobias said to be sure to “keep your eyes open”. A non-animal origin declaration doesn’t mean that you don’t need to thoroughly understand the process of vendors and always apply the precautionary principle. Ulrich said there are so many definitions of non-animal origin, it is important to understand what a supplier means by their definition. Ask the supplier, conduct a thorough audit, and ensure that their definition and process matches with what you had in mind.

To learn more, please see CellPrime® Non-animal Origin Supplements and LONG® R3 IGF-I human

This post is sponsored by the SAFC® portfolio brand of MilliporeSigma