Feliza Mirasol is the science editor for 

Feliza Mirasol

Biopharmaceutical manufacturing largely involves biomolecules whose stability depends on temperature. Antibodies, insulin, and, more recently, messenger RNA (mRNA)-based vaccines are examples of biotherapeutics that require cold storage. This article explores the difficulties with developing stability-enhancing formulations for temperature-sensitive biomolecules and technology innovations that are enabling their formulation. 

Prioritizing Formulation Strategies for Temperature-Sensitive Biotherapeutics

Factors that have been most challenging in developing formulations that enhance stability in temperature-sensitive biomolecules include buffer, pH, and sugar. These are usually the typical factors that need to be evaluated to thermodynamically stabilize monoclonal antibodies (mAbs), notes Joseph Cao, PhD, senior scientist, Analytical and Formulations Development, Bionova Scientific. He explains that there are different types of buffers, sugars, and a range of pH to choose from when developing a proper formulation for mAbs that will prevent aggregation, degradation (such as deamidation, oxidation), and particle formation. Besides those factors, amino acids, salts for ionic strength, and excipients are also important in developing a formulation recipe, he adds.

In addition, the limited amount of therapeutic protein to work with poses another challenge to formulation development. “With design of experiment (DoE), it has allowed us to cut the amount of protein and help us efficiently screen for factors,” Cao says. “Freeze-thaw cycles, container combability, and manufacturing impurities should also be considered during formulation development.”

Heonchang Lim, director, Formulation Development, Samsung Biologics, agrees that finding the ideal range of pH and buffer conditions while selecting the optimal formulation condition to get a good thermal stability property are primary factors that challenge formulation development in these molecules. However, he notes, the thermal stability problem can mostly be solved by using optimal pH buffer and excipient combination.

“Nowadays, we bump into another challenge to develop an optimal formulation condition. That is regarding solubility and hydrophobicity of the molecule. Though clients would prefer higher concentrations, when taking unique characteristics of molecules into consideration, formulation scientists cannot always get high enough concentrations. In addition, pH shift may occur due to the Donnan effect,” Lim says. “Consequently, when a client requests a highly concentrated formulation, particularly above 100 mg/mL, [the] formulation team at Samsung Biologics implements rigorous gate checks to prevent such challenges from arising during formulation stages.”

Meanwhile, Caroline Ailhas, Pharmaceutical Development director, CARBOGEN AMCIS says that, for her, the main factors challenging formulation development for temperature-sensitive molecules revolve around maintaining the structure and functionality of the molecule and defining the required duration of stabilization. The latter can vary from a few days—to allow for manufacturing and visual inspection before storage at 2–8 °C—to several months, or even years. “The definition of this duration significantly impacts the formulation strategy,” Ailhas says.

“One of the key performance indicators for the development of biotherapeutic formulations is colloidal stability to prevent protein aggregation and potentially precipitation,” Greg Chrimes, site head and vice-president, Analytical and Formulation Science, KBI Biopharma, adds. He points out that proteins may denature and aggregate at interfaces such as air/liquid, liquid/liquid, and liquid/solid. Chemical modifications and degradation may also impact effector function, efficacy, conformational stability, and other important parameters, he emphasizes.

Pharmaceutical Technology’s Trends in Formulation

When referring to this article, please cite it as Mirasol, F. Prioritizing Formulation Strategies for Temperature-Sensitive Biotherapeutics. 

Articles

Solving the challenge of better-stabilized temperature-sensitive biomolecules hinges on innovative formulation strategies.

The biopharmaceutical industry has long been working on methods to delivery biologic therapeutics in a manner that will enhance ease-of-use and patient compliance. Biologics today are largely administered parenterally, a method that can be painful and undesirable for patients. Furthermore, parenteral formulations of biologics require a strict temperature-controlled environment. Researchers have long been exploring means to formulate biologic-based molecules that can ensure their molecular stability and mitigate some of the stringent manufacturing requirements. In this episode of the Drug Solutions podcast, Feliza Mirasol, science editor of 

® explores the issues challenging biologic formulation and the progress made so far in moving toward alternative routes of biologic drug delivery with Nigel Theobald, CEO of N4 Pharma, a specialist pharmaceutical company developing novel delivery systems for oncology, gene therapy and vaccines.

Nigel Theobald is CEO of N4 Pharma, a specialist pharmaceutical company developing novel delivery systems for oncology, gene therapy and vaccines. Theobald has over 25 years’ experience in healthcare, in building businesses, strategy development and implementation, and in a strong network covering all aspects of pharmaceutical product development and commercialization. He was the head of healthcare brands at Boots Group plc in 2002 before leaving to set up a series of successful businesses, including Oxford Pharmascience Group plc, which he grew over five years into an AIM-quoted company with a market capitalization of £40 million (US$53 million) upon departure. Theobald formed N4 Pharma in 2014.

 presents the Drug Solutions podcast, where the editors will chat with industry experts from across the pharmaceutical and biopharmaceutical supply chain. Join us as experts share insights into your biggest questions—from the technologies, to strategies, to regulations related to the development and manufacture of drug products.

Podcasts

In this podcast episode, we discuss both the challenges of and the progress made so far by the biopharma industry toward non-parenteral means of delivering biologic drugs.

Drug Solutions Podcast: Alternative Drug Delivery Methods for Biologics: Where Are We At?

Lonza announced on Sept. 24, 2024 that it has entered into a long-term commercial supply agreement with Vertex Pharmaceuticals under which it will manufacture Casgevy (exagamglogene autotemcel), the first clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated protein 9 (Cas9)-gene-edited cell therapy for treating sickle cell disease (SCD) and β thalassemia.

Under the agreement, Vertex will have access to Lonza’s scientific, regulatory, and manufacturing expertise as well as its global manufacturing network and its experience in the commercial manufacture of cell therapy products. Lonza plans to manufacture Casgevy at its cell therapy manufacturing facilities in Geleen, the Netherlands, with plans to expand to its Portsmouth, NH manufacturing facility in the United States.

“It is a privilege to work with Vertex on bringing its innovative and cutting-edge medicines to patients suffering from life-threatening diseases. We are also pleased to reach a significant milestone towards supporting the commercial manufacture of Casgevy by receiving the regulatory approval at our state-of-the-art cell therapy manufacturing site in Geleen,” said Daniel Palmacci, president, Cell & Gene, Lonza, in a company press release.

The Geleen facility recently received a good manufacturing practice (GMP) license from FDA, the European Medicines Agency (EMA), and the UK Medicines and Healthcare products Regulatory Agency (MHRA). Lonza expects the Portsmouth site to begin GMP operations in 2025.

“Manufacturing a first-of-its-kind therapy [such as] Casgevy is complex and requires advanced technology and capabilities. The Lonza team have been excellent partners as we have invested in our global manufacturing network to ensure Casgevy will be available for the patients who need it. We look forward to our continued collaboration,” said Morrey Atkinson, PhD, executive vice-president and chief technical operations officer, head of Biopharmaceutical Sciences and Manufacturing Operations, Vertex, in the press release.

Casgevy was approved by the MHRA in November 2023 (1), by FDA in December 2023 (2), and by EMA in February 2024 (3). Following the MHRA and FDA approvals in late 2023, Vertex 

, a UK-based cell and gene therapy contract development and manufacturing organization, in December 2023 (4) to manufacture Casgevy. RoslinCT runs manufacturing facilities in Edinburgh, Scotland, and Boston, Mass., which are purpose-built for cell therapy products. The CDMO had been working with Vertex on an adaptive basis while Vertex moved through clinical and regulatory development of Casgevy.

1. Vertex Pharmaceuticals. Vertex and CRISPR Therapeutics Announce Authorization of the First CRISPR/Cas9 Gene-Edited Therapy, Casgevy (exagamglogene autotemcel), by the United Kingdom MHRA for the Treatment of Sickle Cell Disease and Transfusion-Dependent Beta Thalassemia. Press Release, Nov. 16, 2023.
2. FDA. 

FDA Approves First Gene Therapies to Treat Patients with Sickle Cell Disease

. Press Release, Dec. 8, 2023.
3. Vertex Pharmaceuticals. European Commission Approves First CRISPR/Cas9 Gene-Edited Therapy, Casgevy (exagamglogene autotemcel), for the Treatment of Sickle Cell Disease and Transfusion-Dependent Beta Thalassemia. Press Release. Feb. 13, 2024.
4. RoslinCT. RoslinCT to Manufacture First-Ever US FDA Approved CRISPR-Based Gene Therapy Casgevy (exagamglogene autotemcel). Press Release. Dec. 11, 2023.

Lonza signed a long-term supply agreement to manufacture CASGEVY (exagamglogene autotemcel) for Vertex at its facility in Geleen, the Netherlands, and plans to expand manufacturing to Portsmouth, NH, in the United States.

Lonza and Vertex Enter Commercial Supply Agreement for CASGEVY, the Sickle Cell Disease Gene-Edited Cell Therapy

With FDA's approved of Eli Lilly and Company’s (Lilly’s) EBGLYSS (lebrikizumab-lbkz), a targeted interleukin 13 (IL-13) inhibitor, a new biologic treatment is made available for moderate-to-severe atopic dermatitis (eczema) in adults and children 12 years of age and older. The treatment is applicable to patients who weigh at least 88 pounds (40 kg) and whose moderate-to-severe atopic dermatitis is not well controlled with topical prescription therapies.

FDA’s approval was based on results from several studies (ADvocate 1, ADvocate 2, and ADhere). These studies included more than 1000 adults and children (aged 12 and older) with moderate-to-severe eczema whose symptoms were not controlled with topical prescription medicines. At 16 weeks, the primary endpoint for these studies was evaluated; measurements showed clear or almost clear skin, according to the company press release.

"Patients still struggle to control their moderate-to-severe atopic dermatitis with currently available therapies. Many experience poor long-term disease control, and severe itch can significantly impact their daily lives," said Jonathan Silverberg, MD, PhD, professor of dermatology at George Washington University School of Medicine and Health Sciences in Washington, DC, and first author of the 

 manuscript summarizing EBGLYSS clinical trials (1), in a company press release. "[FDA’s] approval of EBGLYSS is a big win for patients, as we now have a new first-line biologic treatment option for moderate-to-severe disease when topical prescriptions aren't enough."

Averaging two of the studies (ADvocate 1 and 2), it was shown that 38% of people who underwent the treatment achieved clear or almost-clear skin at 16 weeks, compared to 12% with placebo. Ten percent saw the same results in as early as four weeks. Many responders also experienced itch relief in both studies with the treatment. On average, 43% felt itch relief at 16 weeks, compared to 12% on the placebo. Five percent of people felt relief as early as two weeks.

"Eczema can affect people of all skin tones, ethnicities, genders, and ages. Nearly 16.5 million adults in the [United States] have eczema, with 6.6 million experiencing moderate-to-severe symptoms [such as] itchiness, dry and scaly skin, discoloration, and rashes, which can lead to more scratching that may cause skin to crack and bleed, (2)” said Kristin Belleson, president and CEO of the National Eczema Association, in the release. "The approval of EBGLYSS provides hope and promise for the eczema community and those still seeking lasting relief from disruptive symptoms."

"People living with eczema have symptoms that can appear at the most inopportune times, creating unpredictability and impacting their everyday lives," said Daniel Skovronsky, MD, PhD, chief scientific officer and president of Lilly Research Laboratories, and president, Lilly Immunology, in the release. "Today's approval allows people the opportunity to reimagine life with eczema as EBGLYSS offers a targeted approach to reduce a main cause of eczema inflammation. EBGLYSS provides long-lasting symptom relief with a convenient once-monthly maintenance dose."

EBGLYSS was also approved in Europe by the European Commission in 2023 and in Japan in January 2024. Additional market approvals are expected later this year.

Lilly has exclusive rights for the development and commercialization of EBGLYSS in the US and the rest of the world outside Europe. Almirall, Lilly's partner, has licensed the rights to develop and commercialize the biologic for treating dermatology indications, including eczema, in Europe.

1. Silverberg, J. I.; Guttman-Yassky, E.; Thaçi, D.; et al. Two Phase 3 Trials of Lebrikizumab for Moderate-to-Severe Atopic Dermatitis. 

 (12),1080–1091. DOI: 10.1056/NEJMoa2206714
2. Chiesa Fuxench, Z. C.; Block, J. K.; Boguniewicz, M.; et al. Atopic Dermatitis in America Study: A Cross-Sectional Study Examining the Prevalence and Disease Burden of Atopic Dermatitis in the US Adult Population. 

 (3), 583–590. DOI: 10.1016/j.jid.2018.08.028

The approval of EBGLYSS (lebrikizumab-lbkz) by FDA puts a new biologic treatment for moderate-to-severe atopic dermatitis on the market for patients whose symptoms are not well controlled with topical treatments.

Lilly’s EBGLYSS Offers New First-Line Biologic Treatment for Atopic Dermatitis Following FDA Approval

Eli Lilly and Company is enhancing its global biopharmaceutical production with an added $1 billion expansion of its manufacturing site in Limerick, Ireland. The company announced the increased investment on Sept. 12, 2024. With the new investment, Lilly will increase production of biologic active ingredients at the site, including the biologic active ingredients for Kisunla (donanemab-azbt), the company's therapy for treating early symptomatic Alzheimer's disease 

The company also unveiled its new $800 million facility expansion in Kinsale, Ireland. The Kinsale facility began operations in 2023 and produces Lilly's latest diabetes and obesity treatments in response to market demand, according to a company press release.

The additional $1 billion investment in Limerick, on top of the $800 million investment in Kinsale, is part of the company’s goal to raise its manufacturing footprint. Lilly has committed more than $20 billion since 2020 into building, expanding, and acquiring manufacturing facilities in both the United States and Europe, the company stated in the release.

"These investments will boost the production of some of our medicines, helping millions of people with diabetes, obesity, and Alzheimer's disease live the healthiest lives possible," said Edgardo Hernandez, executive vice-president and president of Lilly Manufacturing Operations, in the release. "We won't stop there—these state-of-the-art facilities will also be equipped to support our promising pipeline molecules of the future."

Once completed, the Limerick site will become a part of Lilly's global manufacturing network for the production of the biologic active ingredients for the company's Alzheimer's disease portfolio as well as other biologics.

"Alzheimer's disease is a devastating diagnosis for both the patient and for their loved ones, along with having a huge burden on society," said Anne E. White, executive vice-president and president of Lilly Neuroscience, in the release. "The treatments we will make here in Limerick offer the potential to be able to slow the progression of the disease and make life better for millions of people around the world."

The new $1 billion investment brings Lilly’s total investment in Limerick to $2 billion, which is double the amount the company initially announced in March 2023. The site will include advanced technologies such as machine learning, artificial intelligence, and automated robotics and systems to enable right-first-time execution. Lilly plans to create another 150 jobs for highly skilled workers in Limerick, including positions for engineers, scientists, quality assurance professionals, and operations personnel, bringing the total staff at the site to 450 employees. Lilly broke ground on the Limerick site in March 2023, following its initial announcement for the new facility in 2022. The company anticipates production of biologic active ingredients to start up in 2026.

Meanwhile, the new Kinsale site will serve the company’s production of diabetes and obesity therapies. The site uses a digital-first process that integrates continuous manufacturing technology for creating a new manufacturing platform for complex peptides.

FDA Approves Treatment for Adults with Alzheimer’s Disease

Lilly has added a $1 billion investment to increase biologics manufacturing at its Limerick, Ireland, site while unveiling its new $800 million facility in Kinsale.

Lilly’s $1.8 Billion Biologics Expansion Expected to Grow Manufacturing Footprint in Ireland

On Sept. 9, 2024, Sartorius announced the launch of its Vivaflow SU tangential flow filtration (TFF) cassette. The Vivaflow SU is designed to be more user-friendly and to allow for more flexibility in laboratory workflows. The new cassette offers more efficient and sustainable ultrafiltration and diafiltration processes for feed volumes in the range of 100 mL–1000 mL.

According to a company press release, the new all-in-one cassette reduces cost and complexity and streamlines workflows for molecule isolation in a sustainable manner. Each cassette is ready to use and has a standard laboratory pump, which enables simple set-up. The simplified set-up avoids the need for expensive systems and expensive process optimization and allows for faster and more reliable concentration and rebuffering of any molecule. The cassette also eliminates the need for cleaning and reduces contamination risk while minimizing waste, according to the press release.

“Vivaflow SU is a testament to our promise to always simplify progress,” said John Cashman, product manager for Lab Ultrafiltration at Sartorius, in the press release. “It takes the ease-of-use that our customers already value about TFF with Vivaflow and elevates the experience with improved handling and performance, while reducing waste. Our next-generation cassette eliminates the need for feed flow rate, TMP [transmembrane pressure], and permeate flux calculations, while also offering sustainability improvements that are truly impressive for a single-use consumable.”

Sustainability features of the new cassette include a new housing that uses 30.5% less plastic. It also incorporates several eco-friendly design improvements, such as locally sourced raw materials, customized tubing kits for each cassette configuration, a digital user manual, and reduced non-recyclable packaging; all these design elements contribute to a lower research footprint, the company stated in the press release.

Vivaflow SU is the-next-generation TFF cassette that replaces the previous Vivaflow 50. It is meant for research scientists who require a fast and reliable means for concentrating and rebuffering large feed volumes in molecule isolation workflows.

The launch of Vivaflow SU follows on the heels of Sartorius’ earlier launch of the Arium Mini Extend water purification system for laboratories in June 2024 (1). The Arium Mini Extend is the newest member of the Arium Mini product line. The new purification system is designed to meet the Type 1 water demands of laboratories for general, life-science, and analytical applications, according to the company in a press release. The new system is compact and offers ultrapure water purification with flexibility and convenience.

The new system comprises a flexible, ergonomic handheld dispenser, a user-friendly digital display, and the Arium Bagtank technology to significantly optimize laboratory workflows. The dispensing unit is equipped with an independent volume control function and expands the access area for ultrapure water. The new system streamlines water purification processes, especially those processes that require repeated dispensing. Meanwhile, operation and maintenance have been made simpler with intuitive menus and a 5-L Bagtank for storage of purified water. These features facilitate cleaning procedures and ensure uninterrupted laboratory workflows, the company stated in its press release.

1. Sartorius. Sartorius Launches New Arium Mini Extend with Flexible Arm for Versatile Type 1 Water Dispensing. Press Release. June 17, 2024.

Sartorius' new Vivaflow SU tangential flow filtration cassette offers better flexibility in lab workflows and is designed to be more user-friendly. 