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Feliza Mirasol

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Certain additives or a combination of additives in downstream bioprocessing have demonstrated an improvement in the separation and clearance of empty and partially full capsids during the manufacturing of viral vectors, noted Ohnmar Khanal, PhD, Downstream Technology Lead, Spark Therapeutics, at the 

American Society of Gene and Cell Therapy’s (ASGCT’s) 27th Annual Meeting

, which is being held from May 7–11, 2024 in Baltimore, Md.

 on the work her team has been doing to test the removal of empty and partially filled adeno-associated virus (AAV) capsids in the downstream processing stage of AAV vector production. Khanal stated that she and her team have been looking at additives and the role they play in AAV purification. In particular, her team focused on copper ions and conducted oversaturated loading experiments using multicolumn anion exchange (AEC) chromatography.

“A current challenge in the downstream purification of recombinant [AAV] is the separation of transgene-containing capsids from empty [capsids] and undesired capsid variants,” Khanal stated. “AEX resin is often used below its saturated binding capacity to separate empty and full AAV particles on a single column. However, the outcome of the traditional single-column operation can be impacted by variability in load composition, sensitivity to buffer composition, and fractionation strategies,” she explained.

Khanal noted that, in reviewing past studies, her team found that the removal of empty and partial capsids with a high yield was rarely demonstrated in the literature. In her talk, Khanal presented three-stage separation strategies for two recombinant AAV constructs (rAAV) and demonstrated a way to overcome the trade-off between yield and purity.

In the first strategy, a resin chemistry (Poros XQ, Poros HS, Poros I) is selected and used with a concoction of kosmotropic agents (e.g., ammonium sulfate or sodium sulfate). These agents (i.e., additives) were used to promote capsid stability at low ionic strength. Meanwhile, metal ions (e.g., copper, magnesium, calcium) were used to promote selectivity of the desired capsid. “Using copper ions improved the resolution between empty and full capsid separation and helped remove other product-related impurities,” Khanal pointed out in her presentation.

In the second strategy, the team used an appropriate resin/media structure and geometry to exclude AAV from smaller resin pores (e.g., Poros XQ). Doing so minimized particle dispersion and improved resolution. “Monolith and membrane chromatography showed better peak resolution than traditional resin-based on favorable transport behavior,” Khanal said.

Finally, in the third strategy, the transition from single column to multiple columns in series was tested. This was done by loading 1.5x the resin saturation capacity. Khanal noted that this allowed for over 40% reduction of partial capsids and over 95% removal of empty capsids. “Using these strategies, we provide the best modality to achieve maximum empty and partial capsid removal with a yield of over 80%,” she concluded.

Articles

Experiments conducted by the downstream technology team at Spark Therapeutics involving metal ion-containing additives showed improved capsid clearance in AAV production.

ASGCT 2024: Additives Play a Role in Removal of Empty and Partial AAV Capsids

US-headquartered ProPharma, a US-headquartered provider of regulatory, clinical, and compliance services for the life sciences industry and a portfolio company of Odyssey Investment Partners, and Italy-based PBL, a custom solutions provider to the pharmaceutical, chemical, and diagnostics industries have launched the Cell Factory Box (CF Box), an enclosed, fully automated cell factory device. The companies launched the device under a partnership. 

The CF Box enables the decentralized manufacturing of all types of cell and gene therapies (CGTs) in Class D (ISO8) or controlled not classified surrounding areas, according to a May 2, 2024 press release.

“Regarding treatment options, many are adopting a decentralized approach to manufacturing [CGTs],” said Eleonora Casucci, vice-president of Quality & Compliance EU, ProPharma, in the press release. “The CF Box provides key advantages compared to the devices currently available. Our collaboration with PBL positions us to serve those in the [CGT] segment fully.”

According to the release, the CF Box uses already-approved artificial intelligence (AI) technology and is extensively automated, allowing it to significantly reduce manual operation. The nature of the device also reduces the need for high-class clean rooms. Using approved AI technology, the CF Box employs batch detection recognition, which is set to a frozen state during validation, and batch manufacturing. Users can select modules based on individual needs and select the type of products/technologies desired. Additional modules can also be added at later stages.

“Robotic arms, cameras, liquid transfer systems, conveyor belts, components, and tools are available in the market, enabling full automation, including but not limited to alerting the operator once the culture has reached the desired values,” the press release states. The design allows for flexibility in automating manual processes and supports diverse manufacturing tasks. The CF Box is also programmable for various processes, including optimized cell culture expansion, and streamlining manufacturing timelines effectively, according to the release.

“We are honored to work with ProPharma on such a monumental effort,” said Filippo Begarani, Research and Innovation manager, PBL, in the press release. “Our first goal is always to conceive, design, and implement technologically advanced solutions with the highest standards of quality and innovation, and the CF Box device will not only change but elevate how [CGTs] are manufactured since it supports high reproducibility of the process at various locations.”

The CF Box will be demonstrated in Frankfurt, Germany, during ACHEMA, which will be held June 10–14, 2024. PBL is also offering demonstrations after the event and upon request at its facility in Parma, Italy.

The global CGT market size is projected to grow to $82.24 billion by 2032 from $15.46 billion in 2022, with a compound annual growth rate of 18.3% (1). Key market drivers are the exponential increase in the CGT clinical pipeline and an increase in the number of regulatory approvals for innovative medicines.

Cell and Gene Therapy Market (By Therapy Type: Cell Therapy and Gene Therapy; By Indication: Cardiovascular Disease, Cancer, Genetic Disorders, Infectious Diseases, Neurological Disorders; By Delivery Method: In-Vivo and Ex-Vivo; By End-Users: Hospitals, Cancer Care Centers, Wound Care Centers, Others)—Global Industry Analysis, Size, Share, Growth, Trends, Regional Outlook, and Forecast 2023-2032

ProPharm and PBL have introduced a fully automated, enclosed cell factory manufacturing device.

New Cell Factory Box Launched by ProPharma and PBL for Automated CGT Manufacturing 

In a new research collaboration and license agreement, Poseida Therapeutics and Astellas Pharma’s wholly owned subsidiary, Xyphos Biosciences,will develop novel 

CAR programs to target solid-tumor cancers.

Under the agreement, the companies plan to combine Poseida’s proprietary allogeneic chimeric antigen receptor T cell (CAR-T) platform with Xyphos’ ACCEL technology to create one Poseida-developed CAR-T construct. This construct is intended to serve as the basis of two 

CAR product candidates targeting solid tumors.

“We are excited to expand our relationship with Astellas, where we share a vision that cutting edge, off-the-shelf cell therapies can address significant unmet needs of patients with solid tumor malignancies. Today’s agreement further reinforces the economic value of Poseida’s highly differentiated non-viral technologies and enables development in areas beyond our core pipeline focus,” said Kristin Yarema, PhD, president and CEO of Poseida, in a May 1, 2024 company press release.

Under the transaction, Xyphos will reimburse Poseida for costs incurred as part of the research agreement. Xyphos will be responsible for the development and future commercialization of products generated from the collaboration. Poseida will receive a $50 million upfront payment as well as potential development and sales milestones and contingency payments that could total up to $550 million. Poseida is also eligible for up to low double digit tiered royalties as a percentage of net sales.

“At Astellas, we have a strong commitment to developing novel treatments for patients with cancer and have positioned immuno-oncology as a [p]rimary [f]ocus of our R&D strategy [1] … we are focused on reinvigorating the immune system’s ability to discover, disarm, and destroy cancers in more patients. By combining the ACCEL platform with Poseida’s elegant and cutting-edge genetic editing platforms, we believe the collaboration will bring synergies between the two companies’ breakthrough research and will ultimately lead to expansion of Astellas’ portfolio and to delivery of innovative CAR-T cell therapies to cancer patients,” said Adam Pearson, chief strategy officer of Astellas, in the press release.

Astellas is making a strategic investment in Poseida, which it announced in August 2023 (2). Under this transaction, Astellas is investing a total of $50 million, comprising $25 million to acquire 8,333,333 shares of Poseida common stock, a one-time $25 million payment for a right of exclusive negotiation and first refusal to license P-MUC1C-ALLO1, one of Poseida's clinical-stage programs. The candidate is an allogeneic CAR-T cell therapy being developed to treat multiple solid tumor indications. Also under this transaction, Astellas has been granted a board observer seat on Poseida’s board. The position includes the ability to attend Poseida’s scientific advisory board meetings as well as certain notice rights related to any potential change of control of Poseida.

 (accessed May 6, 2024).
2. Astellas Pharma. Astellas and Poseida Therapeutics Announce Strategic Investment to Support Poseida’s Commitment to Redefining Cancer Cell Therapy. Press Release, Aug. 8, 2023.

The two companies will combine their cell therapy platforms to develop convertibleCAR programs targeting solid tumors.

Poseida Therapeutics and Astellas to Develop Novel Allogeneic Cell Therapies

Data are the biopharma industry’s most valuable asset today, which is prompting the industry to step up its measures to not only gather these data, but to protect them as well. In a conversation with Nathalie Batoux, product manager, Research Innovation, IDBS, 

 delved into the approaches the biopharma industry has taken to shore up data integrity protection. 

Editor's note: this article was published in the Pharmaceutical Technology Europe May 2024 issue.

 Explain why data are the biopharma industry’s most valuable asset. The analogy is that there’s a gold rush for data (1); what does that mean?

 With the current digital transformation happening in the biopharma industry, the industry has also seen the explosion of [tools such as] artificial intelligence (AI) and machine learning (ML). There is also a significant amount of data that exists now based on the products already on the market. With the explosion of AI and ML, people want to reuse the data that they have. So, instead of running new experiments, already existing data are being used; these historical data are very important. There are some new initiatives underway to review and repurpose existing drugs for other applications, thus, AI and ML are being used to review these already existing data. Not only is the biopharma industry producing a vast amount of new data, it also has its hands on a significant amount of historical data.

 Describe the impact of this perception—this ‘gold rush’ for data—on data integrity. Has it led to a flurry of activity?

Yes, there is a flurry of activity. There are many startups that are proposing AI and ML to the biopharma industry, amongst other industries. There are initiatives, as I mentioned, around the repurposing of existing drugs using historical data at hand.

 Can you discuss where data integrity practices in the biopharma industry are the strongest and where they are the weakest?

 My perspective is to look at the evolution within the industry, and where we are now with digitalization, data integrity is getting much stronger. Looking at the practice of manually recording, or manually transcribing, data from one system to another, this is very much error prone. Whether intentionally or accidentally, errors are highly likely. If there is an error in the data, then any analysis conducted later on will be incorrect, and the decision-making using that data will not be good and will thus be counterproductive. This will cost money at the end of the day.

Good data integrity involves the use of more automated systems, using digital transformation where the system records the context around the results. For example, in an experiment, scientists—even though they are very well trained—are still human. There’s only a finite number of parameters that they can deal with. While a computer, or a series of computers, can record all sorts of things around the context. It’s as simple as the date and time of an experiment that might change the outcome, whether [the experiment] is done in the summer or winter. That is important as temperature and humidity conditions may affect the outcome.

The other advantage is that, with data digitally recorded, there is traceability, and most of the modern systems now have audit capabilities. [This makes it easier] to understand what has happened to generate that data, but also what has happened after the data has been generated—such as, has the data been transferred from one system to another? Having all that context, having this history, ensures that, in the first place, the data—that is, the data that will be used to train the models for ML or analysis, or for use by AI—will be of quality. [And that assurance comes from] all that context, all that history behind the data.

Data that are digitally recorded [are] easier to find. [In the laboratory], finding an experiment on paper or in a notebook is not easy. [Digitally recorded data] offer interoperability and [better] accessibility. If [the data are] on paper, for example, [the lab tech] needs to have that piece of paper in front of [them] to be able to see it. This hinders interoperability [because] that piece of paper, if the handwriting is, as they say, like a doctor’s handwriting, not many people will be able to understand it. Being ‘reusable’ as a whole is very, very important for data, so having FAIR [findable, accessible, interoperable, and reusable] data is one of the principles of data integrity. This will enable the digital transformation and all those initiatives that are happening with ML and AI.

 Let’s talk about the data itself. Good data versus bad data; how is that determined?

 As mentioned earlier, having all the context around the data recorded is one of the attributes of good data. The other attributes, as I mentioned, are the FAIR principles; however, one needs to make sure that they are recording everything. It’s human nature to try to record things that are happening and [what is] working, but from a learning perspective—and for ML, in particular—one needs to also record what didn’t work because [there is] much more to learn from things that didn’t work. So, the ‘how to’ recognize good data versus bad data requires having rigour, using well documented SOPs [standard operating procedures], and having controls in place, all of which are quite important because these help with the reproducibility of that data.

In addition, if using a digital system with controls in place, then [the technician] will receive an alert, and [this allows] them to record an exception. It is very important to record all those things, not only for good data in general, but also for the data that regulatory bodies require. [Keep in mind], all the compliance requirements are very important. Another point [to highlight] is that for accessibility and interoperability, more than one system will need to be used; likely, it will require several systems. And, data needs to be connected easily, so having silos that are not talking to each other, that don’t represent each other, that is not good data. Having the ability to connect the data—to connect to the structure of that data in order to understand them—that is a symptom of good data.

 How is the biopharma industry handling this flow of data; how is it preparing for this gold rush that was mentioned earlier?

 [With the] digitalization that is happening, less companies are using paper, and [it seems that] more and more companies have been investing in data scientists to be able to mine the data. However, there is a difference between having all that data mined by specialists and the people who are actually using the data at the bench, so having democratization of data is extremely important. Empowering [bench] scientists with decision-making based on large data sets, including past data, at the click of a button is very valuable. Rather than having to wait for a few weeks for data scientists’ analyses—although the data scientists’ skills are still important for more complex analyses—a more diverse set of scientists or end users can have accessibility to data with [the goal] of helping to put algorithms in place for the analyses, and so on.

1. Batoux, N.; Rayner, D. In a Digital Gold Rush, Data Integrity is Priceless. 

®
Vol. 36, No. 5
May 2024
Pages: 33–34

When referring to this article, please cite it as Mirasol, F. Shoring Up Data Integrity Practices. 

Digital transformation is allowing for better handling, analysis, and protection of vast data collection.

Shoring Up Data Integrity Practices

Considerations for the Use of Human- and Animal-Derived Materials and Components in the Manufacture of Cell and Gene Therapy and Tissue-Engineered Medical Products

, has been made available for public comment by FDA (1). FDA’s Center for Biologics Evaluation and Research issued the draft document in April 2024.

In the draft guidance, manufacturers of cellular and gene therapy (CGT) and tissue-engineered medical products (TEMPs) are provided recommendations for assuring the safety, quality, and identity of materials—both human and animal origin—that are used to manufacture CGTs and TEMPs. Recommendations are given in regard to the chemistry, manufacturing, and control (CMC) information related to the use of human- and animal-derived materials. This type of information is submitted for an investigational new drug application.

Materials covered by this draft guidance include the reagents, feeder cells, excipients, and other inactive ingredients in the drug product that are in direct contact with the starting material, intermediates, and final products as well as any materials used to manufacture reagents, feeder cells, and excipients, and materials incorporated in TEMPs, according to FDA (1). Materials excluded from the guidance include human cells used as starting material to manufacture human cells, tissues, and cellular and tissue-based products, including TEMPs.

The agency states in the document that “Use of human- and animal-derived materials during product manufacturing may increase risks of infectious disease transmission, and raises potential safety concerns, such as the possible introduction of adventitious agents or other impurities into CGT products and TEMPs. Thus, human- and animal-derived materials should be thoroughly characterized and described in your regulatory submission” (1).

Comments can be submitted electronically or in writing by the deadline date on July 29, 2024, after which time, the agency will begin working on a final version of the guidance (2).

According to the agency, this guidance is intended to supplement the two final industry guidance documents, 

Chemistry, Manufacturing, and Control (CMC) Information for Human Gene Therapy Investigational New Drug Applications (INDs)

Guidance for FDA Reviewers and Sponsors: Content and Review of Chemistry, Manufacturing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs) 

Draft Guidance for Industry, Considerations for the Use of Human-and Animal-Derived Materials in the Manufacture of Cell and Gene Therapy and Tissue-Engineered Medical Products

Considerations for the Use of Human-and Animal-Derived Materials and Components in the Manufacture of Cell and Gene Therapy and Tissue-Engineered Medical Products; Draft Guidance for Industry; Availability

Guidance for Industry, Chemistry, Manufacturing, and Control (CMC) Information for Human Gene Therapy Investigational New Drug Applications (INDs)

. Guidance for FDA Reviewers and Sponsors, Content and Review of Chemistry, Manufacturing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs)

A new draft guidance issued by FDA covers human- and animal-derived materials used in the manufacture of advanced therapy medicinal products.



New FDA Draft Guidance Made Available for Materials Used in Manufacturing CGTs and Tissue-Engineered Therapeutics

FDA's Center for Biologics Evaluation and Research (CBER) has issued a new draft guidance, 

Safety Testing of Human Allogeneic Cells Expanded for Use in Cell-Based Medical Products 

(1). The document was issued in April 2024.

According to FDA in an April 30, 2024 notice in the 

 (2), “Allogeneic cells of human origin may be expanded in culture to manufacture medical products consisting of live cells, inactivated cells, cell lysates, or other cell-based materials such as cell-derived particles.” Thus, the draft guidance provides recommendations to help sponsors of allogeneic cell-based medical products determine what cell safety testing is appropriate to support an investigational new drug application (IND) or a biologics license application (BLA).

The agency notes that cell safety testing should be based on a risk analysis that considers the expansion potential of the cells in addition to the reagents that are used to expand these cells in culture as well as the number of individuals capable of treatment with the cell-based medical product.

Allogeneic cell-based products that are regulated by CBER’s Office of Therapeutic Products are subject to this new guidance. It applies to cultured allogeneic cells—including cell banks—that serve as sources of intended constituents for the final drug product. The guidance also applies to combination products that contain an allogeneic cell or cell-based biologic component in combination with a drug and/or device.

“The recommendations in this guidance also apply to genetically modified allogeneic cells that have been transduced with viral and/or plasmid vectors, and cells that have undergone genome editing. This guidance does not apply to cell substrates that are used during manufacturing of non-cell-based products such as viruses, gene therapy vectors, or recombinant proteins,” the draft guidance document states (1). However, the document emphasizes that recommendations for feeder or other cells used as reagents during manufacturing are beyond the scope of this particular guidance.

FDA is accepting comments on the draft guidance either electronically or in writing until the deadline of July 29, 2024, after which time it will begin work on the final version of the guidance (2).

FDA has stated that this new draft guidance is intended to supplement two other final industry guidance documents, 

Content and Review of Chemistry, Manufacturing, and Control (CMC) Information for Human Somatic Cell Therapy Investigational New Drug Applications (INDs)

Draft Guidance for Industry, Safety Testing of Human Allogeneic Cells Expanded for Use in Cell-Based Medical Products

Safety Testing of Human Allogeneic Cells Expanded for Use in Cell-Based Medical Products; Draft Guidance for Industry; Availability

In the new draft guidance, FDA provides recommendations on cell safety testing of human-sourced allogeneic cells for sponsor companies.