TNKase is the first stroke drug to win FDA approval in nearly three decades.

The FDA signed off Monday on the use of Roche’s thrombolytic drug TNKase to treat acute ischemic stroke in adult patients.

According to Roche subsidiary Genentech, which announced the label expansion on Monday, TNKase is the first new drug for stroke in almost 30 years. Roche and Genentech also own Activase, the only other acute ischemic stroke (AIS) drug approved by the FDA.

Delivered intravenously, TNKase is a tissue plasminogen activator that works by kicking off a cascade that culminates in dissolving blood clots. In AIS, this mechanism helps prevent the formation of blockages that can constrict the flow of blood to various regions of the brain. TNKase was first approved in 2000 to lower the risk of death in patients with acute heart attack.

Monday’s label expansion was backed by data from a large multicenter trial that established the non-inferiority of TNKase to Activase. Data published in July 2022 in The Lancet showed that 36.9% of patients treated with TNKase had no to nonsignificant disability as measured by the modified Rankin Scale, a validated tool that doctors use to assess global disability in stroke patients.

In comparison, 34.8% of comparators in the Activase arm reached the same outcome. The risk difference estimate was 2.1%, which satisfied the threshold of non-inferiority.

In a statement on Monday, Genentech Chief Medical Officer Levi Garraway called TNKase’s label expansion a “significant step forward” for the company and for patients, for whom the drug can provide “a faster and simpler administration, which can be critical for anyone who is dealing with an acute stroke.”

Monday’s label expansion also continues Roche’s regulatory run in recent months.

Last month, for instance, the company’s SMN2 splicing modifier Evrysdi became the first FDA-approved tablet for spinal muscular atrophy, a rare motor disorder. The pill “combines established efficacy with convenience,” Garraway said at the time, and is expected to make the drug easier to take for patients.

Months earlier, in September 2024, Roche—alongside partner Sanofi—won the FDA’s approval for the industry’s first biologic treatment for chronic obstructive pulmonary disease. Dupixent, a blockbuster anti-IL4-alpha antagonist, was cleared for the lung condition after Phase III trials demonstrated a 30% to 34% drop in the rate of exacerbations versus placebo.

In April 2024, Roche’s Genentech also snagged approval for Alecensa as the first and only ALK inhibitor for the adjuvant treatment of ALK-positive non-small cell lung cancer patients with early-stage disease who had undergone surgical resection.

The High Street pharmacy chain Boots is asking customers to return packs of 500-milligram paracetamol tablets because a labelling error incorrectly states they are a different painkiller, aspirin.

More than 110,000 packs, with the batch number 241005 and expiry date “12/2029” on the bottom, are affected.

Customers can receive a full refund without a receipt.

Boots and the supplier, Aspar Pharmaceuticals Limited, have begun a full investigation.

Conservation strategies are turning back the doomsday clock in threatened Florida-Scrub Jays — but not without caveats, a new study published in Current Biology shows.

In the early 2000s, conservationists proposed a plan to move isolated jays to a region comprising thousands of acres of restored habitat, home to a small community of 13 jays.

Translocation, where an organism is moved from one area to another, offers a means to prop up declining populations. Across an eight-year stretch from 2003 to 2010, 51 jays were relocated from fragmented and degraded habitats to a partially restored, contiguous region of scrubland called the M4 Core Region.

This strategy was proposed to thwart the compounding factors putting the Scrub-Jay at risk of extinction: inbreeding, decline in population size and reduced genetic diversity.

A team of researchers led by MSU conservation geneticists Tyler Linderoth and Sarah Fitzpatrick analyzed decades’ worth of data, finding that translocations successfully bolstered population numbers but failed to overcome genetic erosion and inbreeding. The

Decades of systematic tagging, field observations and genetic sequencing provided a nearly complete pedigree of the jays.

Leveraging this rich dataset, the researchers analyzed the genetic consequences of this strategy, sequencing the entire genomes of 87 jays sampled before, and several generations after, the first translocated jays were introduced to the M4 core region.

This study shows, in unparalleled resolution, how demographic mechanisms, including births, deaths, emigrations and immigrations, influenced the genetic conditions of the M4 core region population.

“The only way to really have a pulse on population health is through both demographic and genetic monitoring, which can inform when and what conservation interventions are needed and how to adapt management accordingly,” said Linderoth, the paper’s lead author.

The researchers identified that although the population had rebounded, growing to ten-fold the original population size, genetic erosion persisted.

This population increase is critical for the continued survival of the species.

The dampened genetic diversity uncovered by the researchers is due to an uneven number of offspring produced per family line, a factor called reproductive skew.

Reproductive skew limits the effective population size: the members of a population who produce the next generation. A high effective population helps ensure robust genetic diversity, while low numbers indicate that genetic variation will decrease more rapidly.

A handful of genetic lines tracing back to mostly translocated jays now dominate the genetic make-up of the jay population, dampening genetic diversity. Importantly, however, the authors were able to show with simulations that translocation efforts effectively pumped the breaks on genetic erosion, despite failing to reverse it.

The authors note that translocations likely provided a net benefit to the population.

“Even though translocations did not completely prevent the loss of genetic diversity, they likely slowed the rate at which genetic diversity within the core population was lost and prevented inbreeding from being as high as it would have been otherwise,” Linderoth said.

The authors hope this study informs future conservation projects, highlighting the viability of translocations as a means for supporting at-risk populations.

“Even though translocations did not completely prevent the loss of genetic diversity, they likely slowed the rate at which genetic diversity within the core population was lost and prevented inbreeding from being as high as it would have been otherwise,” Linderoth said.

The authors encourage future projects to anticipate the negative impact of reproductive skew on translocation strategies and stress the importance of habitat management in supporting these efforts.

“Without sound habitat management and protection, translocations are likely doomed to fail. Even small areas of habitat can serve as important stepping-stones that facilitate migration and connectivity between populations,” Fitzpatrick said.

The MSU researchers partnered with ecologists & co-authors Raoul Boughton, from The Mosaic Company, and Lauren Deaner of Flatwoods Consulting.

For over 20 years, ecologists from The Mosaic Company have monitored groups of Florida Scrub-Jays located 25 miles from the state’s west coast, monitoring changes at the demographic and genetic levels.

The conservation project first began with a partnership between The Mosaic Company, Reed Bowman — bird biologist at Archbold Biological Station, and pioneer of a 54-year Scrub-Jay monitoring program — and The United States Fish and Wildlife Service.

Raoul Boughton, lead ecologist at Mosaic and a collaborator on the study, explains the results detailed in this publication stem from a 30-year commitment to monitor and analyze the results of the mitigation translocation.

“This publication highlights the genetic outcomes of this extensive experiment to date and provides critical information on how we may further improve the success of this project,” Boughton said.

Researchers from the University of Toronto have shown how a low-carbohydrate diet can worsen the DNA-damaging effects of some gut microbes to cause colorectal cancer.

The study, published in the journal Nature Microbiology, compared the effects of three different diets — normal, low-carb, or Western-style with high fat and high sugar — in combination with specific gut bacteria on colorectal cancer development in mice.

They found that a unique strain of E. coli bacteria, when paired with a diet low in carbs and soluble fibre, drives the growth of polyps in the colon, which can be a precursor to cancer.

“Colorectal cancer has always been thought of as being caused by a number of different factors including diet, gut microbiome, environment and genetics,” says senior author Alberto Martin, a professor of immunology at U of T’s Temerty Faculty of Medicine.

“Our question was, does diet influence the ability of specific bacteria to cause cancer?”

To answer this question, the researchers, led by postdoctoral fellow Bhupesh Thakur, examined mice that were colonized with one of three bacterial species that had been previously linked to colorectal cancer and fed either a normal, low-carb or Western-style diet.

Only one combination — a low-carb diet paired with a strain of E. coli that produces the DNA-damaging compound colibactin — led to the development of colorectal cancer.

The researchers found that a diet deficient in fibre increased inflammation in the gut and altered the community of microbes that typically reside there, creating an environment that allowed the colibactin-producing E. coli to thrive.

They also showed that the mice fed a low-carb diet had a thinner layer of mucus separating the gut microbes from the colon epithelial cells. The mucus layer acts as a protective shield between the bacteria in the gut and the cells underneath. With a weakened barrier, more colibactin could reach the colon cells to cause genetic damage and drive tumour growth. These effects were especially strong in mice with genetic mutations in the mismatch repair pathway that hindered their ability to fix damaged DNA.

While both Thakur and Martin emphasize the need to confirm these findings in humans, they are also excited about the numerous ways in which their research can be applied to prevent cancer.

Defects in DNA mismatch repair are frequently found in colorectal cancer, which is the fourth most commonly diagnosed cancer in Canada. An estimated 15 per cent of these tumours having mutations in mismatch repair genes. Mutations in these genes also underlie Lynch syndrome, a genetic condition that significantly increases a person’s risk of developing certain cancers, including colorectal cancer.

“Can we identify which Lynch syndrome patients harbour these colibactin-producing microbes?” asks Martin. He notes that for these individuals, their findings suggest that avoiding a low-carb diet or taking a specific antibiotic treatment to get rid of the colibactin-producing bacteria could help reduce their risk of colorectal cancer.

Martin points out that a strain of E. coli called Nissle, which is commonly found in probiotics, also produces colibactin. Ongoing work in his lab is exploring whether long-term use of this probiotic is safe for people with Lynch syndrome or those who are on a low-carb diet.

Thakur is keen to follow up on an interesting result from their study showing that the addition of soluble fibre to the low-carb diet led to lower levels of the cancer-causing E. coli, less DNA damage and fewer tumours.

“We supplemented fibre and saw that it reduced the effects of the low-carb diet,” he says. “Now we are trying to find out which fibre sources are more beneficial, and which are less beneficial.”

To do this, Thakur and Martin are teaming up with Heather Armstrong, a researcher at the University of Alberta, to test whether supplementation with a soluble fibre called inulin can reduce colibactin-producing E. coli and improve gut health in high-risk individuals, like people with inflammatory bowel disease.

“Our study highlights the potential dangers associated with long-term use of a low-carb, low-fibre diet, which is a common weight-reducing diet,” says Martin.

“More work is needed but we hope that it at least raises awareness.”

Continuous processing can get products to market about 12 months faster than batch processing, according to a 2022 paper by the FDA. Understandably, the drive to transition to continuous bioprocessing is strong.

For fermentation products, the bulk of the transitional work has involved E. coli platforms. Little has been done to speed the transition for another industrial-scale expression platform, Bacillus licheniformis, which is known for its rapid growth, high productivity, and high-purity output.

Julian Kopp, PhD, a postdoctoral researcher at Technical University Vienna (TU Wien), and colleagues there and at White Biotechnology Research assessed the transferability of B. licheniformis from batch and fed-batch fermentation to continuous cultivation.

“The literature states fed-batches are applicable for transitioning to chemostat cultivation for other inductive systems,” Kopp, the paper’s corresponding author, tells GEN. “It was a surprise for us, therefore, to see that chemostat cultivations have to be screened individually from fed-batch cultivations, and that researchers have to establish hard data with continuous cultivations in order to determine suitable operating ranges.”

In a recent paper, they report significant differences in productivity between fed-batch and chemostat cultivations, despite similar growth rates. Kopp and colleagues found continuous systems showed “a clear dependence on generation time.” To ensure stable productivity for recombinant protease production, those cultivations needed dilution rates between 24% and 50% and specific substrate uptake rates above 30%. Fed-batch cultivations, conversely, needed low growth rates to enhance productivity.

Biopharmaceutical developers aiming to transfer B. licheniformis from fed-batch to continuous cultivation processes, they conclude, should not rely upon the data from fed-batch screenings.

A closer look at their data shows that chemostat cultivations with an intermediate dilatation rate of approximately 40% of the maximum uptake rate were optimal for product formulation. Productivity dropped significantly both below 25% and above 72%. Biomass formation remained consistent, although productivity, measured in terms of protein concentration, declined over time.

“Fed-batch cultivations showed a strong effect of uptake rate in regard to biomass formation and specific protease productivity,” the researchers note. That system, however, showed a 30% increase of space-time yield when compared to continuous cultivation. “We attributed this effect to the higher amount of energy being applied, whilst still maintaining derepressed conditions, in comparison to the constant-fed reference processes,” they report.

Throughout their experiments, even the worst of the fed-batch cultivations significantly outperformed continuous cultivations. For perspective, the least productive screening had a yield of 80, specific protease productivity of 119 (+/- 31), and an uptake rate of 48 (+/-1). Meanwhile, the most effective continuous cultivation screening had a space-time yield of 46 (+/- 11), a specific protease productivity of 69 (+/- 17), and an uptake rate of 50 (+/-4).

“Our results indicate that in a derepressed induction system, a narrow operational range that supports either biomass growth or induction is essential for effective continuous cultivation,” the researchers conclude.

New research from scientists at Cleveland Clinic’s Genome Center and their collaborators at other institutions describes a pathway that human herpes simplex-1 (HSV-1) can use to contribute to the development of Alzheimer’s disease. They have also identified two FDA-approved drugs that successfully reversed the pathway in the lab. Full details are published in Alzheimer’s & Dementia: The Journal of the Alzheimer’s Association in a paper titled, “Human herpesvirus-associated transposable element activation in human aging brains with Alzheimer’s disease.”

Feixiong Cheng, PhD, Genome Center director and senior author on the study, claims that their findings provide concrete evidence of a possible link between human herpesviruses and Alzheimer’s disease. Many people around the world are either currently infected or will contract herpesviruses by adulthood. Some infections are asymptomatic while others cause minor illnesses. However, even after the illnesses subside, infected individuals carry the virus for the rest of their lives.

While herpesviruses are generally harmless when they are suppressed, there is evidence that shows that immune systems can lose the ability to suppress them under certain conditions including as people age. Circumstantial evidence from other studies has linked HSV-1 to Alzheimer’s disease, but the exact mechanism was not known.

Cheng and his team hypothesized that latent HPV-1 infections could trigger Alzheimer’s disease by directly activating the transposable elements that they had previously connected to disease progression in aging brains. Transposable elements are small pieces of DNA that can physically “jump” out of chromosomes and randomly move to far-away regions of our DNA.

For this study, the researchers mapped all of the transposable elements associated with Alzheimer’s disease in aging brains. The investigators then analyzed four publicly available RNA sequencing datasets from hundreds of healthy and Alzheimer’s-affected brain cells.

They identified several transposable elements that were more highly activated in Alzheimer’s-affected brains containing HSV RNA, compared to uninfected or healthy brains. They then tested HSV-1 infected brain cells to see whether the identified transposable elements were activated. They also assessed any effects on neuroinflammation and protein accumulation, which are associated with Alzheimer’s disease.

What they found was that when people either contract HSV-1 or when latent infections become active with age, the infection is linked with the activation of transposable elements like LINE-1. Once these are activated, they disrupt important genetic processes in the brain that are associated with the accumulation of Tau and other Alzheimer’s-linked proteins which contribute to inflammation and neurodegeneration in the brain.

Next, the scientists analyzed publicly available health records to see if people who were prescribed antiviral herpes medications were less likely to be diagnosed with Alzheimer’s later in life. They found evidence suggesting that two herpes medications, valacyclovir and acyclovir, were associated with significantly reduced instances of Alzheimer’s disease. When they tested the drugs in virus-infected human brain organoid models, they seemed to successfully reverse the activation of transposable elements that impact the Alzheimer’s disease pathway.

Commenting on the study, Cheng noted that their findings could open a door to “new strategies for treating other neurological diseases associated with herpesviruses or other viruses.”

Caliway Biopharmaceuticals has recently announced the completion of its initial public offering (IPO) and up-listing from the Emerging Stock Market to the Taipei Exchange (TWSE-6919). The round raised approximately $206 million (NT$6.4 billion), marking it the largest IPO in Taiwan’s biotech industry history and valuing the company at nearly $3 billion. The company is poised for a transformation in 2025, advancing its groundbreaking clinical programmes and strengthening its global market presence. Following its recent record-breaking IPO, BioSpectrum Asia took an opportunity to speak with Vivian Ling, Chief Executive Officer & Chief R&D Officer, Caliway Biopharmaceuticals to explore their innovative contributions in biopharmaceuticals.

Which products are currently under development?

2025 will be a defining year for Caliway as we push ahead with key clinical advancements and corporate milestones, bringing CBL-514 closer to market. CBL-514, a first-in-class small-molecule drug designed to selectively induce adipocyte apoptosis, provides a non-invasive alternative to liposuction for non-surgical fat reduction in medical aesthetics. We are preparing to secure IND approvals for two pivotal Phase 3 studies from the US FDA and Health Canada, a critical step in advancing CBL-514 as the world’s first investigational drug for large-area localised fat reduction. Beyond fat reduction, we’re also submitting a Phase 2 IND application for a new indication focused on improving body weight rebound, expanding CBL-514’s potential applications.

In Q1, Caliway announced positive Phase 2b study results for CBL-514 (0205 Study), the second and final Phase 2b before moving into Phase 3. In Q2, we are preparing for regulatory discussions with the FDA (EOP2) and EMA to align on the next steps for CBL-514’s late-stage development.

A key focus in Q4 will be completing patient enrollment for the Phase 2b study of CBL-514 in Dercum’s Disease (CBL-514 0202DD). CBL-0202DD is being developed as a potential first-in-class therapy for Dercum’s Disease, a rare and painful condition, and has already been granted Fast Track Designation by the FDA and Orphan Drug Designation by both the FDA and EMA. In the early-stage pipeline, CBA-539 offers a novel approach to hyperpigmentation and skin ageing by inhibiting melanin production and transmission, reducing dark spots and evening skin tone, while also stimulating collagen production to improve skin elasticity and firmness for natural, long-lasting results. Expanding into therapeutic applications, CBL-0201OB targets post-weight loss fat accumulation in combination with GLP-1, with a Phase 2 IND submission planned for Q4 2025.

On the corporate front, we are moving forward with a stock split to enhance market liquidity and investor engagement. Additionally, Caliway is now part of the MSCI World Small Cap Index, originally a Q4 goal further strengthening our global investor presence.

In October 2024, we completed our IPO, raising $206 million, making it the largest IPO in Taiwan’s biotech industry history and positioning us among the top biotech IPOs in the US in 2024. This strong financial foundation provides the necessary resources to advance CBL-514 into global pivotal Phase 3 studies, explore additional indications, and further expand our pipeline.

How do you plan to utilise the funds raised from your recent IPO to advance R&D? 

We are strategically deploying IPO funds to accelerate clinical development, expand global partnerships, and strengthen commercialisation efforts. Our key areas of investment include CBL-514 Phase 3 Studies. A significant portion of the funds is being directed toward launching pivotal global multi-centre Phase 3 studies for CBL-514, ensuring a smooth regulatory pathway in key markets. Focusing on new indications development, we are expanding CBL-514’s applications beyond fat reduction by advancing studies for Dercum’s Disease and weight rebound prevention, broadening its potential.

To leverage international licensing and investment experts, we are actively engaging with global investment and licensing professionals to expedite partnership negotiations, increase visibility among potential collaborators, and secure the most favourable commercialisation deals.

For clinical collaboration with KOLs and investigators, we are strengthening relationships with renowned clinical researchers to expand study participation, enhance scientific credibility, and increase visibility in international markets.

We are also expanding global business development and market positioning; and engaging global pharmaceutical companies for licensing and partnerships. We are actively negotiating potential global licensing agreements discussions and strategic partnerships to drive CBL-514’s commercial success.

We are further strengthening our presence at key global industry events to connect with strategic partners and investors by participating in global industry conferences.

Most recently, we participated in IMCAS 2025 in Paris, where we presented the advancements in clinical progress. These efforts maximise our growth potential, drive regulatory approvals, and ensure long-term commercial success.

Could  you tell us about strategic partnerships or licensing agreements with other pharmaceutical companies, key to accelerating the development and commercialisation of your products?

Strategic partnerships are a key driver of Caliway’s growth. We are actively engaging with leading pharmaceutical companies for licensing and co-development opportunities to accelerate CBL-514’s commercialisation. We are also deepening industry connections through key industry events, including BIO, IMCAS, AMWC, JP Morgan Healthcare Conference, and the World Orphan Drug Congress, ensuring we stay at the forefront of global biotech and aesthetic medicine collaborations. These partnerships will be crucial in accelerating product commercialisation and maximising CBL-514’s global impact.

What are your strategies for expanding the market reach of CBL-514, especially in regions like Taiwan, China, Korea, and Southeast Asia?

We are executing a multi-faceted market entry strategy to ensure a structured and phased approach to regulatory approvals and market commercialisation. CBL-514 is a 505(b)(1) first-in-class small-molecule drug designed to address an unmet need in non-surgical fat reduction. Given its innovative mechanism of action and strong clinical data, our primary entry strategy is to focus first on regulatory approvals in the US, our key reference market. Once established, we will gradually expand into additional key regions, including Asia. We are actively engaging with global pharmaceutical companies for potential licensing and other strategic partnerships to accelerate commercialisation. Our pivotal global Phase 3 studies will further strengthen CBL-514’s market valuation and licensing potential, paving the way for successful entry into international markets.

Focusing on the Taiwan biotech market in particular, what are the existing challenges and opportunities?

Biotech development comes with its challenges. Regulatory processes can be complex, which may impact drug development timelines. Another key challenge is the shortage of specialised R&D talent, particularly in pharmaceutical sciences and clinical research. That said, Taiwan has significant advantages that make it an attractive hub for biotech innovation. Taiwan’s healthcare system is highly advanced, cost-effective, and well-structured. The high density of hospitals, cutting-edge medical technology, and experienced medical professionals make it an ideal environment for research, especially in niche indications and rare diseases. Taiwan’s market agility and adaptability also set it apart. The biotech sector here moves fast, making it easier to pivot and innovate. Additionally, many global pharma companies use Taiwan as a strategic entry point, launching products before expanding into larger markets. With its strong medical ecosystem, advanced infrastructure, and strategic position in the region, Taiwan offers a unique and competitive environment for biotech growth and drug development.

Thermo Fisher Scientific agreed to acquire Solventum’s purification and filtration (PF) business for approximately $4.1 billion in cash. The PF business provides purification and filtration technologies used in the production of biologics as well as in medical technologies and industrial applications. The Solventum business operates globally with sites across the Americas, Europe, the Middle East, Africa, and the Asia-Pacific region, and has approximately 2,500 colleagues. In 2024, Solventum’s PF business generated approximately $1 billion in revenue.

Solventum’s PF business is highly complementary to Thermo Fisher’s bioproduction business, according to Marc N. Casper, chairman, president, and CEO of Thermo Fisher. Thermo Fisher has a portfolio of offerings in cell culture media and single-use technologies and Solventum’s filtration portfolio broadens Thermo Fisher’s capabilities in the development and manufacturing of biologics, spanning upstream and downstream workflows, said Casper.

“The addition of Solventum’s business is an outstanding strategic fit with our company and will create significant value for our customers and shareholders,” added Casper. “Solventum’s portfolio of solutions will be valued by our customers, and further demonstrate our disciplined capital deployment strategy which has an excellent track record of creating shareholder value.

“As the trusted partner to our customers, Solventum’s PF business will expand and add differentiated capabilities to our bioprocessing portfolio to better serve our customers in this rapidly growing market.”

The transaction is expected to be completed by the end of 2025 and is subject to customary closing conditions and regulatory approvals. Once the transaction closes, Solventum’s PF business will become part of Thermo Fisher’s Life Sciences Solutions segment.

As new targeted biologic modalities ramp up for commercialization, manufacturing methods must keep pace. If new efficacious therapeutics cannot be produced at reasonable costs they remain in restricted use or are shelved for economic reasons in lieu of other options.

For example, autologous CAR T-cell therapies have demonstrated remarkable success, but manufacturing costs remain extremely high due to the large footprint and substantial clean room costs. Miniaturizing manufacturing and bringing it to the point-of-care setting could, potentially, benefit more patients at a lower cost to the healthcare system.

CAR T cells are just one example. Many processes that produce recombinant proteins use either mammalian or microbial cells and could reap cost savings if development and optimization at small scale better reflected that at scale-up or scale-out volumes. A promising resource, microbioreactors are just beginning to show their might as they become more sophisticated with integrated sensors and automation instrumentation.

A microbioreactor is a way of “doing more with less” succinctly stated Wei-Xiang Sin, PhD, research scientist at SMART CAMP. As these tiny growth machines become even more sophisticated with machine learning and AI algorithms they can only positively impact the future of biologics production.

Point-of-care manufacturing

Over a decade ago, Kevin Lee, PhD, and Harry Lee, PhD, and their colleagues in the MIT laboratory of Professor Rajeev Ram, PhD, developed the 2 mL “Breez” microbioreactor platform technology, which was subsequently spun out as Erbi Biosystems. In 2020 Millipore-Sigma, the U.S. and Canada Life Science business of Merck KGaA, acquired Erbi Biosystems and expanded their Mobius bioreactor portfolio.

“We have been working with the Breez since its development,” said Sin. Various designs of the microfluidic chip, ranging from 100 µL to 1 and 2 mL working volumes, have been used for microbial and mammalian cell culture applications.

“We thought that automated, closed-system microfluidic bioreactors might also be a novel production approach for the personalized nature of autologous cell therapies,” said Sin. Autologous cell therapy manufacturing has lengthy processes, with large equipment footprints, low production throughputs, the need for centralized cleanroom facilities, and a high cost of goods. “In particular, the perfusion-capable, modular Breez can support extremely high viable cell densities in a small volume and footprint,” continued Sin. The parallelized format, with four “pods” per system, allows up to four simultaneous runs per system to raise production throughputs and enable efficient scale out.

The 2 mL design was used to test human CAR T-cell production. Various timelines of activation and transduction as well as two different perfusion schemes, were evaluated to determine optimal conditions.¹ Minimal system modifications were made in this proof-of-concept study. Engineering improvements should help move the Breez closer to GMP compatibility on the way to potentially enabling decentralized, point-of-care manufacturing.

The small working volume could reduce the amount of GMP-grade viral vectors and reagents and thus the costs associated with CAR T-cell manufacturing. Importantly, the Breez has the smallest footprint (0.044 m2 per dose) compared to existing manufacturing methods, noted Sin. This attribute substantially decreases cleanroom fixed costs.

“The ability to make clinical-scale CAR T-cell doses in an extremely small form factor essentially means doing more with less—more production runs in parallel with less reagents, space, and manpower,” said Sin.

Better prediction and optimization

Many biological drugs are produced using mammalian cells. The process begins with cell line development (CLD) to determine which cell lines will produce the highest levels of recombinant proteins while maintaining stability during large-scale manufacturing.

According to Cheng-Han (Charles) Tsai, PhD, CEO at Cytena BPS, in the CLD workflow cell lines undergo incremental scaling of culture from static formats to shaker flask expansion and, eventually, to bioreactors. However, static formats lacking agitation technology face significant limitations. The size of multi-well plates and the cell numbers often restrict the ability to effectively agitate the culture, which in turn limits oxygen transfer and the overall culture environment. Microbioreactors offer small-scale, controlled culture environments that allow for better oxygen transfer and optimized cell growth conditions that can replicate the conditions needed for larger-scale bioreactors.

Currently focused on CLD applications, Cytena BPS’s microbioreactors provide precise control over culture conditions, enabling faster and more efficient cell line screening and optimization, according to Tsai, who adds that “additional applications include spheroid culture, stem cell/iPSC culture, long-term proliferation, metabolism and dynamic cellular behavior monitoring.”

The company’s C.NEST^® microplate agitation culture system is designed for high-throughput screening in 96- and 24-well plate formats. Customizable mixing intensities allow adjustment of the agitation levels according to the specific cell type and concentration improving oxygen transfer and environmental conditions. In addition, the S.NEST™ system incorporates sensors that provide real-time measurement of dissolved oxygen (DO) and pH to provide more accurate assessments of cell growth conditions to accelerate cell line development, improve process optimization, and efficiently evaluate cell culture health.

“Customers report that introducing mixing early has accelerated their scale-up processes, increased cell concentrations at each stage, and reduced the number of passaging steps,” said Tsai. Notably, the Cytena BPS workflow not only reduced a client’s CLD process time but also significantly increased cell viability in later-stage selection. “Facilitating tests at smaller scales while allowing for precise control of production parameters can permit better prediction and optimization of results before scaling up to larger production volumes,” he added.

Expediting screening

When contemplating the addition of microbioreactors to a workflow, Cristina Martija-Harris, product manager, Beckman Coulter Life Sciences, recommends evaluating usability, scalability, reproducibility, and reliability, as well as compatibility with existing data management and analysis tools. Suppliers can assist with a cost-benefit analysis to determine the economic viability of adoption.

The automated high-throughput BioLector XT Microbioreactor expedites the screening process for different microbial strains/clones, explained Martija-Harris. Applications are diverse including  food and beverage, microbiome studies, agriculture, and many aspects of academic, pharmaceutical, and biotech R&D.

The microbial screening platform allows users to design and execute sophisticated experiments that align with biological signals, enhancing scalability and reproducibility, Martija-Harris continued. Online measurements increase data reliability and robustness. In combination with the Biomek i5 Liquid Handler workstation the system permits individually triggered actions such as sampling, dosing of inducers or feed solutions, and inoculation of culture wells in a microtiter plate. “These actions are executed in response to real-time signals from the microbioreactor, including biomass, pH value, DO concentration, and experiment time without interruption to the shaking of the microtiter plate,” said Martija-Harris.

She pointed out that the BioLector XT Microbioreactor allows efficient clone selection along with problem solving during process development and optimization when there are many mutually influencing parameters.² The system utilizes a standard 48-well microtiter plate format that operates with online, pre-calibrated optical sensors for real-time measurement of cultivation parameters. Patented microfluidic technology facilitates concurrent pH control and feeding processes per cultivation well.

An optional Light Array Module (LAM) provides customizable light settings of 400-700 nm within the photosynthetic spectrum. Sixteen different LED-types can be controlled individually to deliver maximum irradiances and photon flux densities up to 3500 µmol/m2/s to support work with light-dependent organisms that require photosynthesis to grow, said Martija-Harris. “One of the standout features of the BioLector XT system is its strict anaerobic module, which is specifically designed to cultivate microorganisms that require an oxygen-free environment,” she explained.