Feliza Mirasol is the science editor for 

Feliza Mirasol

Sandoz, the generics and biosimilars division of Novartis, announced on March 9, 2023 that it has signed a memorandum of understanding to build a new biologics production plant in Lendava, Slovenia. The company expects to invest at least $400 million in this project, which represents one of the largest international private-sector investments in Slovenia, according to a company press release.

Work on the new plant is set to begin in 2023, and full operations are provisionally planned for late 2026. The facility will support the increasing global demand for biosimilars.

“Biosimilar medicines increase access to cutting-edge biologic therapies for the patients who need them most. This investment underscores our ambition to be the sustainable global leader in biosimilars, a segment projected to grow double-digit annually over the next decade,” said Richard Saynor, CEO, Sandoz, in the press release.

“This state-of-the-art site will … enable[e] us to meet growing demand for our current and future biosimilars in the mid- to long-term. The location offers us a strong combination of political stability, proximity to our existing European-based production and commercial operations, and competitive costs,” said Glenn Gerecke, global head of Technical Operations, Sandoz, in the release.

In addition to this biologics production facility, Sandoz also 

recently announced in 2022 an additional €50 million (US$53 million) investment

 (1) to expand its European-based antibiotics production network. Altogether, the company’s total new investment commitment in the past few years to expand its European-based antibiotics production network comes to €250 million (US$266 million).

1. Sandoz, “Sandoz Announces Further Investment in Key Manufacturing Facility in Austria, to Support Increased Global Demand for Essential Antibiotics,” Press Release, Nov. 7, 2022.

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Sandoz, a Novartis division, is expected to invest at least $400 million to build a new biologics manufacturing facility in Lendava, Slovenia, to support growing demand for biosimilars.

Sandoz to Build New Biologics Production Facility in Slovenia

On March 16, 2023, Catalent announced that it has signed a licensing agreement with India-based Bhami Research Laboratory (BRL) that will provide Catalent with access to BRL’s formulation technology. Using the technology, Catalent intends to develop formulations that will help enable the subcutaneous delivery of high-concentration biologics. Financial terms of the agreement were not disclosed.

The agreement authorizes Catalent to collaborate with its customers to evaluate BRL’s formulation technology so that Catalent and its customers can explore ways to reduce viscosity and deliver high-concentration biologic products. BRL’s technology is applicable to a wide range of monoclonal antibodies and fusion proteins. Successful programs could be integrated into Catalent’s formulation and manufacturing services at scale.

According to a Catalent press release, the subcutaneous delivery of small volume, low viscosity biotherapeutics can provide superior patient benefits over traditional intravenous injections. Such benefits include improved access to healthcare, reduced need for hospitalization and overall treatment cost, and the opportunity to employ new delivery technologies, such as auto-injectors.

“The patented, high-concentration, low-viscosity protein drug delivery platform has been designed to make patient care convenient, affordable, and safe,” said Bhami Shenoy, co-founder and chief scientific officer of BRL, in the press release.

“This agreement allows Catalent Biologics to offer its customers a novel patented formulation that may be evaluated for enabling the delivery of high-concentration therapies by reducing viscosity,” said Julien Meissonnier, Catalent’s chief scientific officer, in the release. “The potential for maintaining the stability of a drug with the convenience of a pre-filled syringe or auto-injector could have a significant impact on reducing the barriers to subcutaneous delivery of therapies and improving the patient experience.”

Under a technology licensing agreement with Bhami Research Laboratory, Catalent intends to develop formulations that will allow for the delivery of high-concentration biologics subcutaneously.

Catalent Licenses Bhami Research Laboratory Technology for Biologics Formulation Development

Genuv, a South Korean-based clinical-stage biotechnology company, announced on March 15, 2023 that it has entered into a partnership with South Korea-based biopharmaceutical company Celltrion to jointly discover and develop novel therapeutic antibodies. Under the agreement, the companies will use Genuv's proprietary SHINE MOUSE platform.

According to the agreement, Genuv will conduct antibody discovery services using its mouse platform technology. Drug discovery will be followed by a research collaboration. Celltrion has the option for joint research and development projects that, if exercised, will trigger agreed-upon milestones, which include development milestones of up to $25 million followed by total commercial milestones of up to $680 million. The latter amount will be triggered dependent upon the accumulation of $7.5 billion in sales per candidate antibody.

The advantage that the SHINE MOUSE platform offers rests in the platform’s ability to generate a greater diversity of antibodies than conventional mice can generate. This ability led to the discovery of the experimental, novel anti-programmed cell death protein 1 monoclonal antibody, GNUV201. GNUV201 holds unique features, such as exceptional interspecies cross-reactivity and binding affinity, which allows for preclinical disease animal model studies that have increased clinical relevance, according to a company press release.

Genuv is also developing GNUV205, an improved interleukin-2-based immunocytokine anticancer drug that is made tumor-specific by leveraging the properties of GNUV201.

“We are excited to embark on our mouse platform business with Celltrion as our first partner,” said Sungho Han, founder and CEO of Genuv, in the press release. “We believe Celltrion's in-depth knowledge and experiences in antibody development and production and Genuv's expertise in novel antibody discovery with our SHINE MOUSE platform could generate great synergies in providing innovative antibody therapeutics.”

“Celltrion is continuously interested in antibody-based new drugs and anticipates to discover differentiated antibodies through innovative antibody discovery platform technology. We look forward to partnering with Genuv for the development of innovative antibody therapeutics that will bring new hope to patients with high unmet needs,” said Kisung Kwon, head of the R&D unit of Celltrion, in the release.

The collaboration between Genuv and Celltrion will utilize Genuv's proprietary mouse platform for antibody discovery.

Genuv and Celltrion Partner on Development of Novel Therapeutic Antibodies

On March 10, 2023, FDA approved US-based Acadia Pharmaceuticals’ Daybue (trofinetide), an oral solution for treating a rare genetic disease. According to an agency press release, the drug is the first treatment for Rett syndrome, a rare, genetic neurological disorder. The drug is indicated for treating adult patients and children two years of age or older who have the condition.

Rett syndrome affects the way the brain develops, and patients with this disorder experience a progressive loss of motor skills and language. A hallmark symptom of Rett syndrome is near constant repetitive hand movements, such as rubbing or clapping. The disorder, which primarily affects females (1 in 10,000) and even more rarely affects males, leads to severe impairments that affect nearly every aspect of life, including speaking, walking, eating, and breathing.

Daybue’s efficacy and safety were evaluated in a randomized, double-blind, placebo-controlled, 12-week study (Study 1; NCT04181723) involving patients with Rett syndrome at five to 20 years of age. Patients treated with the drug demonstrated a statistically significant difference compared to placebo on co-primary efficacy endpoints, according to the agency press release.

The drug is expected to be available in the United States by the end of April 2023, according to an Acadia Pharmaceuticals company press release. Acadia entered into an exclusive license agreement with Neuren Pharmaceuticals in 2018 for the development and commercialization of trofinetide in North America for the Rett syndrome indication as well as other indications.

“Rett syndrome is a profoundly debilitating and complex, rare, neurodevelopmental disorder that presents differently across patients and can lead to an array of unpredictable symptoms throughout the course of a patient’s life,” said Jeffrey L. Neul, Annette Schaffer Eskind chair and director, Vanderbilt Kennedy Center, professor of Pediatrics, Division of Neurology, Pharmacology, and Special Education, Vanderbilt University Medical Center, and Phase III study (LAVENDER) investigator, in the Acadia Pharmaceuticals press release. “Now, for the first time after decades of clinical research, healthcare providers finally have a treatment option to address a range of core behavioral, communication, and physical symptoms for their patients living with Rett syndrome.”

FDA has approved Acadia Pharmaceuticlas’ Daybue (trofinetide), the first treatment for Rett syndrome, a rare genetic disorder.

Acadia Pharmaceuticals Receives FDA Approval for First Treatment for Rare Genetic Disorder

On March 10, 2023, Pfizer announced that FDA approved ZAVZPRET (zavegepant), which, according to a company press release, is the first and only calcitonin gene-related peptide receptor antagonist nasal spray for migraine. The drug is approved for the acute treatment in adults of migraine with or without aura.

FDA’s approval is based on two pivotal randomized, double-blind, placebo-controlled Phase III studies that established the drug’s efficacy, tolerability, and safety profiles. The results showed that ZAVZPRET was statistically superior to placebo on the co-primary endpoints of pain freedom and freedom from most bothersome symptom at two hours post-dose. Study results also demonstrated that pain relief was achieved as early as 15 minutes in a prespecified secondary endpoint versus placebo. Following approval, the drug is expected to be available in pharmacies in July 2023.

“The FDA approval of ZAVZPRET marks a significant breakthrough for people with migraine who need freedom from pain and prefer alternative options to oral medications,” said Angela Hwang, chief commercial officer, president, Global Biopharmaceuticals Business, Pfizer, in the press release.

“Among my migraine patients, one of the most important attributes of an acute treatment option is how quickly it works,” said Kathleen Mullin, associate medical director at New England Institute for Neurology & Headache. “As a nasal spray with rapid drug absorption, ZAVZPRET offers an alternative treatment option for people who need pain relief or cannot take oral medications due to nausea or vomiting, so they can get back to normal function quickly.”

FDA has approved Pfizer’s ZAVZPRET (zavegepant), a new nasal spray treatment for migraine. 

FDA Approves Pfizer’s New Migraine Nasal Spray

Bioassays play a crucial role in biologic drug development as a means to determine potency, toxicity effects, pharmacological activity, stability, purity, and other product qualities. The complexity of biological molecules, however, requires a wide range of analytical procedures to adequately characterize the product. In particular, the concern over immunogenicity in the use of biologics is driving the need to develop more highly sensitive assays. Because of this concern, there is a continual need to refine biological assays to increase accuracy and reproducibility. There is a specific need to replace 

 bioassays are important tools employed during drug discovery/drug development, according to Weihong Wang, PhD, director of Technology Development, Eurofins BioPharma Product Testing. 

 bioassays use cell culture and offer the benefit of low cost and high throughput, both of which are essential during early development. An 

 bioassay also has the necessary robustness needed for use as a lot-release assay for product quality control later in the drug development life cycle, Wang explains.

 bioassay is a more efficient method that allows researchers to clearly characterize product mechanism of action in a controlled environment, says Catherine Liloia, director, Cell & Microbiology, PPD clinical research, Thermo Fisher Scientific. “This approach is scalable and reproducible across production and can assess product stability,” she states.

 bioassays (non-clinical) use animal models and are considered more relevant to potential clinical outcome, says Wang. 

 bioassays are often costly, however, and are less robust and subjected to strict regulations and compliance standards. “Regulatory agencies across the world have been encouraging sponsors to replace, reduce, and refine (the three Rs) dependence on animal studies by advancing development of, and evaluating new, fit-for-purpose methodologies,” Wang explains.

 bioassays are more representative of the full biological complexity and pharmacokinetics of the human body; they can, therefore, more broadly assess potential patient safety impacts stemming from metabolic changes and other side effects. The downside, however, is that 

 bioassays require high investment, strict animal care protocols, and are not as conducive to high-level analytics, according to Liloia.

, generate valuable data needed for regulatory filings, such as investigational new drug (IND) applications. 

 bioassays support determination of product potency, for instance, says Liloia. “When included in analytical panels during release and stability testing, potency data can demonstrate lot-to-lot consistency and long-term stability of the product,” she says.

In addition, bioassays provide data to better understand the impact of manufacturing process changes through process development. “Early 

 methods may be needed to help discern true product mechanism of action in biological pathways that are complex or not well characterized. These methods can sometimes be consolidated or replaced on a product release panel as data [are] collected to determine a single method’s capability to reliably determine product potency across production samples,” Liloia explains.

 method cannot by itself model the biological response in a reliable manner, as is the case with immunogenicity and other safety testing, Liloia adds.

IND-enabling studies, including pharmacology/pharmacodynamic, pharmacokinetics, and toxicology studies, are typically done through both 

 assays, states Wang. She adds that additional 

 assays to address product safety and quality, such as viral testing, and potency testing, are also necessary later on for product licensing.

Cell-based bioassays have also become an important tool in the analytics arsenal for biologics drug development. According to Wang, cell-based bioassays measure relevant physiological responses in cell cultures, but unlike 

 bioassays, cell-based bioassays do not involve the use of live animals and often have different types of assay readouts/endpoints. “Nevertheless, the physiological responses are reflective of mechanism of action, and, therefore, cell-based assays are considered necessary components of product characterization, in addition to other biophysical/physiochemical analytical methods,” she states.

Cell-based bioassays utilize biologically relevant cell lines to measure product potency, says Benjamin Ziehr, senior group leader, PPD clinical research, Thermo Fisher Scientific. He notes that there are several types of cell-based bioassays that vary depending on what is being measured. “The classic cell-based bioassay compares the response of cells treated with drug sample or reference standard. Typically, sample and standard are diluted in a manner suitable to produce a four-parameter response curve in the target cell line. Well-defined response curves are critical for maintaining and demonstrating control of the assay and drug product in these assays,” he explains.

Ziehr further explains that the relative activity of sample drug material to reference standard drug material is reported as the relative potency of the drug product. In comparing cell-based bioassays to other types of bioassays, the key distinction lays in the fact that cell-based bioassays allow mechanism-of-action-dependent measurements, whereas other bioassays measure physical characteristics of the drug product, such as epitope binding or receptor occupancy. “Therefore, cell-based bioassays allow measurement of the cellular processes responsible for a drug product’s effects on the patient,” says Ziehr.

Meanwhile, there are several other modes of cell-based bioassays that provide information about other attributes of the drug product. Ziehr points out two common examples of these other modes from cell and gene therapy product testing: infectivity and replication competency assays. “These assays measure the infectious potential of the drug product and the presence of replication-competent viral particles, respectively,” he states.

In other words, rather than measuring a cellular response, these infectivity and replication competency assays measure the accumulation of viral genetic material following infection of permissive cell lines. “While these assays don’t measure product potency, they do provide critical process and safety data and detect lot-to-lot variability of products,” says Ziehr.

Cell-based bioassays become a clear front runner in this movement, where applicable, remarks Wang, who points out that, in other circumstances, sponsors may be able to eliminate certain 

 assays based on risk analysis. For example, per the International Council for Harmonisation Q5A(R2) (2), 

 testing is no longer necessary for well-characterized cell lines such as Chinese hamster ovary, non-secreting murine myeloma, and SP2/0 (a standard mouse myeloma cell line), based on cell-line history, prior knowledge, and other risk-based considerations, notes Wang.

Despite their advantages, however, cell-based bioassays are limited in their ability to represent more complex, distal biological responses in a patient, cautions Liloia.

Meanwhile, the challenges in working with cell-based bioassays include not only the fickleness of working with certain cell lines, but also the fact that the day-to-day variability is much higher in terms of assay performance, observes Ziehr. The state in which cells enter the bioassay affects their response in the bioassay.

“Cellular state is dictated by many factors, notably metabolic stimuli, cellular density—both in culture leading up to a bioassay and in the bioassay itself—and cellular age. Stable transfections, optimized banking, and robust maintenance controls are needed to minimize cell variability,” Ziehr says.

Ziehr explains that, typically, robust maintenance controls involve the use of well-defined and controlled culture protocols, qualification of critical reagents, and substantial system suitability criteria in the bioassay. “Consistency of technique is challenging, even with clear procedure, so robust training with clear connection between biological processes and cell handling technique remains important,” he remarks.

Because cell-based assays utilize live cells, they are subjected to intrinsic assay variability that are often larger than that of physiochemical methods, says Wang. Thus, the selection of an appropriate cell line that is not only relevant to product mechanism of action, but also able to support a robust response in a dose dependent manner, is the single most critical factor for successful method development, she emphasizes.

“Many details need to be considered and optimized to produce a robust assay suitable for quality control. In addition, for products that may have multiple mechanisms of action, a carefully selected matrix containing multiple cell-based assays may be necessary to fully characterize the product,” Wang says.

Wang also points out, however, that it may yet be difficult to develop a cell-based bioassay for some products that either have not known defined mechanisms of action or have complex mechanisms of action. Furthermore, some cell-based assays still require extended testing time, which can be a cause for concern regarding products that require rapid factory-to-patient delivery. In such cases, Wang suggests that development of surrogate assays and/or orthogonal assays may be beneficial, though subject to discussions with the relevant regulatory agency.

In terms of advantages conferred, compared to 

 bioassays, cell-based bioassays often offer the advantages of a more streamlined testing procedure, higher assay throughput, quicker turnaround time, and less assay variability, according to Wang. “It is generally easier to develop a cell-based assay with an accurate quantitative endpoint than an 

 assay, which is particularly essential when the method is intended to be used for quality control,” she states.

Liloia notes that cell-based methods are quite challenging in many areas. Solutions such as liquid handling automation helps in providing accurate and precise sample dilution pipetting and transfers to the low concentrations and small volumes often needed in a cell-based assay. She also notes, however, that analytical workflows have become more complex because of the diversity of analytical readouts needed to support expanding product areas.

The growing complexity of analytical workflows has resulted in a greater need for coordination across functional areas, broadened cross-training, and design changes to laboratory spaces to maintain efficiency while also maintaining process flows that consider biosafety and contamination prevention, Liloia says. “Data analysis expectations continue to evolve as laboratories and agencies gain experience with newer products, requiring strong statistical support and the ability to apply industry best practices in a new way,” she states.

Overall, cell-based bioassays are considered a critical tool to predict quality and safety of drug products in the patient. “When established early in the development process, cell-based bioassays can provide early indicators of lot-to-lot variability in manufacturing, drug potency stability, and impact from process changes,” states Ziehr.

“Bioassays have an essential role throughout the life cycle of product development of biologics.Many aspects of product discovery, development, characterization, and quality control require careful construction and implementation of bioassays in order to thoroughly assess product safety, efficacy, and stability,” Wang adds.

1. Mire-Sluis, A.R. Progress in the Use of Biological Assays During the Development of Biotechnology Products. 

, 18, 1239–1246. DOI: 10.1023/A:1013067424248.
2. ICH, Q5A(R2) 

Viral Safety Evaluation of Biotechnology Products Derived from Cell Lines of Human or Animal Origin

As the development of increasingly complex biologics grows, more robust bioassays are required to accurately characterize these molecules.