Over the past two decades, 32 cell and gene therapies (CGTs) have been approved by the regulatory authorities worldwide, and more than 2000 novel products are under clinical investigation (1). However, many patients do not have access to these life-saving therapies, partly due to the complexity and high cost of manufacture (2). But how can CGTs be made more cost-efficient given the high labor and facility costs?

According to Miguel Forte, CEO, of Kiji Therapeutics, France, and president of the International Society for Cell & Gene Therapy, “It is clear that CGTs have the opportunity to deliver enormous value to patients; nonetheless, manufacturing is complex, and it has been challenging to deliver the product. Innovative technologies and services have enabled the industry to move towards an automated, closed-system approach that is easier and safer to operate and is less costly and closer to the patient.”

 from autologous cell therapies (patient to patient), allogenic cell therapies (donor to patient), or 

 cell therapies (cell line to patient). Each approach requires manufacturers to tailor the biomanufacturing process to generate specific cell types (and/or vectors), safely, efficiently, and cost-effectively. Manufacturing tools have tended to lag behind scientific advances in CGTs leading to bottlenecks in cell production. Currently, most CGT manufacturers use disconnected single-step/modular or semi-automated solutions; however, the industry is migrating towards fully automated cell therapy manufacturing processes (3). And this approach could significantly reduce manual intervention, risk of contamination, and human error; improve product quality and speed of delivery; and reduce production costs (4).

According to David Smith, vice president of Development, BioCentriq, “Labor costs, and facility costs are sky high, which have led to high price points, particularly in autologous cell therapies where production is 1:1. Automation is a key part of improving costs, reducing labor both in and outside the clean room, and implementing release by exception can help to reduce bottlenecks. Companies are trying to integrate unit operations whether it’s an ‘all-in-one-box’ approach or a ‘unit-operator’ approach where units can be plugged in or out and are integrated within robotic systems to offer greater flexibility.”

For instance, Lonza’s Cocoon Platform and Miltenyi Biotec CliniMACS Prodigy offer an ‘all-in-one-box’ approach providing a closed, automated system for patient-scale cell therapy manufacturing from cell isolation, activation, transduction/transfection, expansion, and harvest within a validated cell manufacturing system (5,6). CliniMACS Prodigy modular Instruments can also be run in parallel to provide large-scale batch production.

UK-based Ori Biotech launched its IRO platform developed in 2024 in collaboration with CTMC (a joint venture between Resilience and MD Anderson Cancer Center). This platform automates the activation, transduction, expansion, and harvest of a wide range of cell types to rapidly scale up manufacturing from the clinic to the commercial setting (7).

Paris-based Astraveus SAS has developed the Lakhesys Benchtop Cell Factory, an end-to-end solution for the manufacture of adherent, non-adherent, and inducible pluripotent stem cells (iPSCs) (8). The company has raised €16.5 million in seed financing and received a €10.4 million grant from the French government as part of its “Innovation in Biotherapies and Bioproduction” initiative, to advance the development of its technology (9).

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Cheryl Barton, PhD is director of PharmaVision.

Article details
BioPharm International®
eBook: Emerging Therapies
September 2024
Pages: 4–7

When referring to this article, please cite it as Barton, C. Frontrunners in Automated Cell and Gene Therapy Manufacturing. 

BioPharm International Emerging Therapies

Articles

Innovative solutions are making personalized cell and gene therapies accessible to all.

Frontrunners in Automated Cell and Gene Therapy Manufacturing

Cell line development (CLD) plays a pivotal role in the production of biotherapeutic molecules. The primary goal during CLD is to generate and identify the most stable cell lines capable of robust production of high-quality therapeutic molecules. To achieve this, screening for both product quality and productivity is essential. Identifying and optimizing stable high-yield cell lines early contributes to accelerating the entire drug development process and the success of the pipeline. 

In addition to classical biologic drugs such as monoclonal antibodies (mAbs), new therapeutic modalities are being developed at a rapid pace. These include bi-, tri-, and multi-specific antibodies, antibody-drug conjugates (ADCs), and fusion proteins such as bispecific T-cell engagers (BiTEs) and peptide-Fc proteins or peptibodies (1,2). The increasing complexity and diversity of these new modalities require innovative analytical approaches that integrate technologies such as capillary electrophoresis (CE) and mass spectrometry (MS). Integrated CE–MS technologies provide the necessary sensitivity, accuracy, and precision to monitor key product quality attributes (PQAs), detect data inconsistencies, and thus characterize, screen, and select optimal cell line clones early in the CLD process.

Given the vast number of potential clones entering the development pipeline, methods to rapidly prioritize candidates are crucial. These methods must be able to quickly and precisely assess the critical quality attributes (CQAs) of the protein products to narrow down the best candidates. Chemical post-translational modifications (PTMs), such as glycosylation, deamidation, and oxidation, are considered CQAs as they often result in charge variants that affect the product’s stability, efficacy, potency, and safety. As such, PTMs must be characterized and controlled throughout the drug development pipeline.

Traditional methods such as enzyme-linked immunosorbent assays (ELISAs) have limitations in sensitivity, specificity, and coverage, which can delay the identification of inconsistencies. Identifying and mitigating data inconsistencies early in the development process is paramount during CLD to ensure the stability and efficacy of the final biotherapeutic product. The integration of CE- and MS-based technologies offers a robust solution that can significantly enhance the detection of data inconsistencies, facilitating the development of stable cell lines for bioproduction.

CE provides rapid assays that can assess key factors affecting final product stability, such as charge and glycan profiles, while MS offers precise information about key factors affecting product quality, such as molecular modifications and their localization and protein structure.

During the manufacturing of mAbs and other complex therapeutics, heterogeneity can arise from enzymatic cleavage and PTMs. Ensuring stability and consistency in therapeutic protein production necessitates meticulous monitoring and control of PTMs and genetic mutations. Moreover, the early detection and characterization of these PTMs during CLD can prevent costly and time-consuming issues later in the production process.

The final therapeutic product can be affected by changes induced by various cell processing and growth conditions, which, therefore, must be comprehensively understood. Alterations in titer, cell culture media, and other process conditions can induce PTMs that affect the stability and activity of the final protein product. Therefore, gathering comprehensive information about key quality attributes, including PTMs, is crucial to understanding the stability of proteins produced by cell lines.

1. SCIEX. Improving the mAb Characterization Process. Whitepaper. Feb. 8, 2024.
2. Zhang, Z.; McCloskey, R. Untangling the Identity of Charge Variants in Antibody-Drug Conjugates. 

Roxana McCloskey is senior global marketing manager, Protein Therapeutics, and Zoe Zhang is manager, Biopharma Applications; both at SCIEX.

When referring to this article, please cite it as McCloskey, R; Zhang, Z. Ensuring Cell Line Stability for Protein Therapeutics. 

Upstream Processing

Stable cell line development can benefit from applying integrated analytical technologies.

Ensuring Cell Line Stability for Protein Therapeutics

Over the past several decades, bioconjugate therapeutics, which unite synthetic chemistry with biological mechanisms, have gained immense interest in research and drug development. Antibody-drug conjugates (ADCs) represent the most prominent class within this modality. Fifteen ADC therapeutics have been approved by FDA since 2000, and more than 150 clinical trials are currently active, roughly 12% of which are late-phase (1). Comprised of a monoclonal antibody (mAb) and a cytotoxic drug compound joined by a linker molecule, ADCs are promising as a novel cancer treatment strategy that enables targeted delivery of potent chemotherapeutic agents while sparing healthy cells and reducing systemic toxicity. However, challenges with off-target toxicities of early ADC candidates slowed the expansion of the field and underscored the importance of linker design in drug safety and efficacy. It is now well-established that linker chemistry and bioconjugation strategies play fundamental roles in the controlled delivery of cytotoxic drugs, presenting new complexities as researchers and drug developers work to optimize ADC performance and deploy linker technology at clinical and commercial scale. 

Unraveling the intricate role of linker design in ADC performance

The function of a linker extends far beyond simply attaching a drug and antibody. It is well-known that the stability of an ADC and the mechanism of payload release can impact ADC performance, and most drug developers recognize that the choice of cleavable or non-cleavable triggers is one of the first decisions to consider. Cleavable linkers employ a chemical or enzymatic trigger within their structure that enables efficient cleavage for controlled drug release within the tumor target. Conversely, non-cleavable linkers do not incorporate a trigger but instead rely on internalization by endocytosis and lysosomal processing to release the payload (2). This choice has a direct impact on the efficacy and toxicity of an ADC, but many other decisions are vital to shaping the final product’s characteristics. The choice of conjugation chemistry, linker backbone, number of payloads per linker, and linker architecture also impact ADC properties such as drug-to-antibody ratio (DAR), ADC aggregation, yield, blood clearance, and the ability of the antibody to bind to its target (3). All of these linker elements can be modified to fine-tune the characteristics of the ultimate ADC product, improving safety, efficacy, stability, manufacturability, and more.

Additionally, linker design can be leveraged to overcome chemical and pharmacological limitations posed by some drug payloads. For example, many payloads are hydrophobic, potentially resulting in poor conjugation yield, aggregation, and increased clearance. Because altering the payload itself to improve hydrophilicity can compromise its activity, linker design can instead be leveraged to modulate ADC solubility and reduce these effects (4). Including a linker with a hydrophilic backbone such as polyethylene glycol (PEG) or modifying the linker architecture to include one or more hydrophilic arms without changing the proximity of the payload to the antibody are options that can effectively shield the hydrophobic payload from the environment, improving conjugation yield and preventing aggregation. Importantly, the variable parameters of ADC components are not independent of one another and must be carefully modified and assessed to maximize desirable features without compromising other functional traits.

1. Sasso, J.M.; Tenchov, R.; Bird, R.; et al. The Evolving Landscape of Antibody–Drug Conjugates: In Depth Analysis of Recent Research Progress. 

 (11), 1951-2000. DOI: 10.1021/acs.bioconjchem.3c00374
2. Bargh, J. D.; Isidro-Llobet, A.; Parker, J. S.; Spring, D. R. Cleavable Linkers in Antibody-Drug Conjugates. 

 (16), 4361-4374. DOI: 10.1039/c8cs00676h
3. Kostova, V.; Désos, P.; Starck, J. B.; Kotschy, A. The Chemistry Behind ADCs. 

 (5), 442. DOI: 10.3390/ph14050442
4. Tsuchikama, K.; An, Z. Recent Advances in Conjugation and Linker Chemistry. 

 (1), 33–46. DOI: 10.1007/s13238-016-0323-0

Pamela James, PhD, is vice-president, Product, at Vector Laboratories.

When referring to this article, please cite it as James, P. Valuing Specialized Experience in ADC Development. 

Manufacturing

Bringing promising ADC candidates to market faster hinges on the biopharma industry investing in linker design capabilities.

Valuing Specialized Experience in ADC Development

When entering the discovery and early development phases of a medication, a company needs to look beyond simply detecting a disease or ailment that could benefit from a novel therapy. There are several factors that should be considered when developing a new modality.

One of these considerations is manufacturing, according to Wade Macedone, CEO of Andelyn Biosciences. “Translating a potential treatment from the laboratory to a therapeutic requires deep knowledge of manufacturing science and technology (MS&T) as well as process development, analytical development, quality and regulatory disciplines,” says Macedone. Viral vector delivery of gene therapies, for example, have manufacturing requirements that are different from antibodies and other modalities. “Everything from the delivery vector, targeting specificity of the serotype, the size of the therapeutic payload and dosing, to the particulars of upstream cell culture and downstream purification will vary from therapy to therapy,” says Macedone. “This is why expertise across the manufacturing disciplines is so important.”

There are two types of cell therapies, explains Michael Pinaud, senior vice president, Business Operations, Ascend Advanced Therapies; autologous cell therapies use patient cells and allogeneic cell therapies use donor cells. Autologous products have more specific logistical challenges. “Material is first collected at an apheresis center, then sent to a central manufacturing location, processed, and finally back to the patient for administration. All this movement takes time and cost and requires maintenance of the chain of identity to ensure the right product gets to the right patient,” says Pinaud. Allogeneic, in comparison, have fewer steps but are costly to produce.

“For either type of cell therapy, costs will be driven by indication, dose required,

 therapy, etc.,” says Pinaud. “Development costs are also very much dependent on the number of patients required for clinical development and the dose.”

Gene therapies, which are also complex, bring in a new or modified gene, turn off a gene, or replace a gene, according to Pinaud. “Most are delivered using viral vectors, which are costly to produce due to challenges with scalability, productivity, and overall recoveries. Limited availability of some raw materials (e.g., plasmids for transient transfection) is also an issue. Fortunately, progress is being made along many fronts, with contract development and manufacturing organizations (CDMOs) taking a lot of the lead in investing in development of optimized and flexible platform manufacturing processes and stable producer cell lines.”

A new cell or gene therapy may require many years and high costs, according to Macedone, with cost varying from indication to indication. Often rare diseases also impact children, which might involve fundraising and patient foundations, Macedone points out, which may aid in costs. “There is always a balance, and every program has its idiosyncrasies. This is where the power of a well-designed modular platform comes into play,” says Macedone.

When referring to this article, please cite it as Haigney, S. The Logistics of Developing an Emerging Therapy. 

Development

Emerging therapies, such as cell and gene therapies, come with unique challenges that must be planned for in advance.

The Logistics of Developing an Emerging Therapy

From cancer to COVID-19, the middle third of 2024 saw a slew of approvals and expanded indications for new and emerging therapies worldwide. Drugs were steadily updated or introduced, and moves were also being made in outsourcing, manufacturing, acquisitions, and new partnerships. The following is a survey of the most impactful developments.

With the fifth anniversary of the COVID-19 pandemic approaching, and as a summer surge increased pressure to respond, FDA on Aug. 22 announced it granted an Emergency Use Authorization for updated versions of messenger RNA (mRNA) SARS-CoV-2 vaccines made by Moderna and Pfizer (1). The vaccines are designed to primarily target the KP.2 strain of the dominant Omicron variant; however, it is likely that the major vaccine developers are already working on a formula for 2025–2026.

The COVID vaccines elevated mRNA into the mainstream lexicon, and while not foreshadowing this new round of shots, when 

® spoke with Christian Cobaugh, PhD, CEO and founder of Vernal Biosciences, at BIO 2024 in San Diego, Calif., in June, Cobaugh addressed the landscape for the outsourcing of mRNA production in 2025 and beyond (2).

“There [are] those that just generally do GMP [good manufacturing practice] for multi different types of drug products, and then there [are] more specialty companies that were set up to make nucleic acids,” Cobaugh said in reference to mRNA service providers, pivoting to a discussion of next-generation sequencing (NGS). “NGS offers … an opportunity to go deeper and pick off multiple critical quality attributes from the same test, which, along with consolidating platforms more broadly, presents significant cost of goods savings in the long run.”

While not an emerging therapy per se, MediWound said on Aug. 15 that NexoBrid, a topically administered enzymatic therapeutic for removal of nonviable burn tissue, was granted a pediatric indication by FDA for treating deep partial and/or full-thickness thermal burns (3).

Treatments for cancer and other diseases of concern

New developments in oncology made headlines in summer 2024. FDA granted a Breakthrough Therapy designation to GSK for an antibody-drug conjugate (ADC) that targets a protein, B7-H3, considered to be a valuable target for cancer therapy (4). The investigational ADC has been under evaluation for treatment of patients with extensive-stage small-cell lung cancer.

Bristol Myers Squibb received validation of a TypeII variation application from the European Medicines Agency for an expansion of the indication for Breyanzi to include adults with relapsed or refractory follicular lymphoma. And the human epidermal growth factor receptor 2 (HER2)-directed ADC Enhertu, developed jointly by AstraZeneca and Daiichi Sankyo, got conditional approval from the National Medical Products Administration of China for treatment of adults with locally advanced or metastatic HER2-positive gastric or gastroesophageal junction adenocarcinomas (5–6).

When referring to this article, please cite it as Lavery, P. Update on Trends in Emerging Therapies and Manufacturing. 

The latest developments in emerging therapies, including new innovations and collaborative efforts in manufacturing.

BioPharm International, Emerging Therapies eBook September 2024

Frontrunners in Automated Cell and Gene Therapy Manufacturing

Ensuring Cell Line Stability for Protein Therapeutics

Valuing Specialized Experience in ADC Development

The Logistics of Developing an Emerging Therapy

Update on Trends in Emerging Therapies and Manufacturing