Continued development of new and next-generation antibodies is spurred by the increasing demand for more effective and custom-tailored biologic therapies. While the bio/pharma industry is seeing a shift toward personalized medicines, ongoing work in research and drug discovery is generating new biologic modalities, such as next-generation antibodies.

In particular, bispecific antibodies are next in line after the decades of success that monoclonal antibodies (mAbs) have had in the industry. Bispecifics were mainly developed for cancer treatment, notes Luca Varani, PhD, Structural Biology group leader at Switzerland’s Institute for Research in Biomedicine, and founder of CLBiotech, a 2022 start-up company focused on nanobody engineering.

Bispecifics (and beyond that, multispecific antibodies) can be simply viewed as a cocktail in a single molecule, or rather, the concept is that of a single molecule that has the property of a cocktail drug. However, Varani emphasizes, bispecifics can do things that a conventional cocktail drug cannot, and that is, bispecifics act synergistically. This is considered a novel mechanism of action, he notes.

Pulling Out All the Stops in mAb Manufacturing

June 1, 2023 BioPharm International® issue

“Bispecifics can achieve molecular functions that are not achievable by monoclonal antibodies,” Varani says. “If you think strategically, you could design a bispecific that acts like a cocktail.”

Ideally, Varani, says, a developer would want to look for bispecifics that do something that monoclonals cannot do, such as, for instance, having a bispecific that bridges two molecules on different cells. Or, another example could be using bispecifics to bridge different viruses, which would cause aggregation with the end result being that the virus is neutralized. Yet again, bispecifics can be used to bridge immune cells, such as T cells, to cancer cells.

“You attach the cancer cell on one end, and you bring the T cell immune system against the cancer with the other,” Varani demonstrates. “You cannot do that with a cocktail.”

Bispecific antibodies present challenges in downstream processing due to “shuffling.” To demonstrate this phenomenon, Varani says to imagine the antibodies having hands. In the case of a mAb, there are two hands. In the case of a bispecific or multispecific antibody, there are four or more hands. Shuffling occurs when the “left hand” of antibody 1, for example, binds with the left hand or the right hand of antibody 2. “This is a mess,” Varani asserts. “This complicates production purification, and you thus have to remove the forms that you do not want.”

“On COVID, for instance, I worked on bispecific [production]. It’s a human format. It’s identical to a monoclonal, but it is bispecific [in function]. Bispecific production yield was very low. Although the cost of production of bispecifics has gone down [significantly], there are still regulatory costs,” Varani says.

“Do you really have a bispecific? Are you producing a molecule that hits both its targets simultaneously? Does it remain a bispecific or does it open up once inside the body, how stable is it? These are all questions for which molecule characterization must be done,” Varani adds.

It is much more difficult to remove unwanted forms due to shuffling in bispecific antibody production compared with mAb production. Because shuffling must be avoided, product yield consequently tends to be lower, Varani says.

Although the purification process has since been improved, Varani notes that some trace amounts of unwanted bispecific antibody forms may be left behind. To that end, the more effective solution to minimizing shuffling and the formation of unwanted product is in the upstream, with molecular engineering, or rather, antibody engineering.

There are a number of solutions that can be derived through engineering. For example, one can engineer what’s known as “single chains” wherein a bispecific antibody can be forced into a single-chain format (rather than a folded molecule, the natural state of an antibody). “This is something very elegant,” Varani says. “You change the interface between heavy and light chains between the two hands [of the antibody] so that they are complementary to each other.”

Looking beyond bispecific antibodies, a new subcategory of antibodies is rising through the ranks: single variable domain on a heavy chain (VHH) antibodies, otherwise known as nanobodies (1), which are derived from camelids (e.g., llamas, alpacas, camels, etc.). One advantage that VHH antibodies hold is that they are single-chain antibodies, making them less structurally complex and smaller in size than human double-chain antibodies. Being single chain, VHH antibodies are not subject to shuffling, making their production much easier, according to Varani. Their small size and simpler structure also make them extremely stable.

In discussing the potential of VHH antibodies to act as immunotherapeutic agents, Varani says: “Being smaller, they have a much higher penetration in tumors. They get in much better than [conventional] antibodies. They can pass biological barriers as well, such as the blood-brain barrier.” Passing through the blood-brain barrier is a task that conventional antibodies cannot do. So, Varani explains, if conventional antibodies are injected into the blood, less than 1% of them will reach the brain, making them ineffective in treating neurological disorders, for instance. The supposition, then, is that VHH antibodies may have the ability to reach targets that conventional antibodies cannot reach.

However, with the pros come the cons. One major disadvantage that VHH antibodies have is that they have a much shorter half-life in vivo, Varani points out. Therefore, duration in the patient’s body may not be sufficient to confer full therapeutic benefit. “They have some advantages, but they also have some problems,” Varani states.

Yet, in the search for next-generation antibodies, VHH antibodies still hold strong potential. According to Varani, development work for VHH antibodies is currently booming after seeing flat activity prior to 2020, and clinical trial programs are in the pipeline. Importantly, the first VHH antibody product, Nanozora (anti-tumor necrosis factor alpha Nanobody), was approved in Japan in September 2022 for treating rheumatoid arthritis (2). Taisho Pharmaceutical developed Nanozora in Japan under a license agreement with Ablynx, a Sanofi company (2).

1. Bever C. S.; Dong J. X.; Vasylieva N.; et al. VHH Antibodies: Emerging Reagents for the Analysis of Environmental Chemicals. 

 (22), 5985–6002. DOI: 10.1007/s00216-016-9585-x
2. Taisho Pharmaceutical, Notification of Approval to Manufacture and Market Nanozora 30mg Syringes for S.C. Injection, a Therapy for Rheumatoid Arthritis, in Japan Japan’s First Nanobody Therapeutic. Press Release, Sept. 26, 2022.

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Features

Exploring new fields can bring novel antibody candidates to the pipeline.

Pushing Forward in Next-Gen Antibody Development

On July 03, 2023, UK-based synthetic biology company Camena Bioscience announced that it has closed a $10 million Series A financing round, which was led by Mercia, a UK-based venture capital firm. With the new funding, Camena intends to scale operations and continue developing gSynth, its enzyme-based DNA synthesis platform.

According to a company press release, demand for DNA synthesis technology has been growing as synthetic genes are being widely used in research to better understand the mechanisms of disease and in the drug discovery process to combat disease. Cost, time, and accuracy of traditional DNA synthesis methods (developed in the 1980s) remains prohibitive, however, particularly when creating long or complex sequences of genes. The technology developed by Camena overcomes these limitations with the use of a novel enzymatic technology that can accurately produce synthetic DNA, according to the press release.

“We are delighted to have closed this latest round of funding with Mercia, which provides further validation of Camena’s approach. The ability to read, write, and manipulate DNA is the foundation of the synthetic biology value chain, yet developments in DNA synthesis have not kept pace with DNA sequencing and editing capabilities,” said Steve Harvey, CEO, Camena, in the release. “Camena’s gSynth platform is revolutionizing the DNA synthesis market by unlocking access to genes and accelerating discovery pipelines.”

“The invention of next-generation DNA sequencing technologies transformed our ability to ‘read’ DNA. Our goal is to enable our customers to ‘write’ DNA with the same confidence and ease,” Harvey added.

“The market for synthetic genes is growing rapidly, but much of the demand is for complex genes which are extremely difficult to produce accurately. Camena’s technology is a game changer that could unlock a much bigger global market. We’re excited to support Camena on its next stage of growth,” said Lee Lindley, investment manager, Mercia, in the release.

New Technology

Camena Bioscience has completed $10 million in Series A financing on the back of growing demand for DNA synthesis.

Camena Bioscience Raises $10 Million to Pursue DNA Synthesis Technology

Thermo Fisher Scientific announced the launch of the Gibco OncePro Tumoroid Culture Medium Kit on June 28, 2023. According to a company press release, the kit is the first commercially available culture medium specifically developed for the expansion of patient-derived tumoroids, or cancer organoids, from multiple cancer indications. In an effort to improve ease of use, the kit includes a scalable, automation-compatible suspension culture method.

According to the release, patient-derived tumoroids hold promise in helping improve clinical trial success rates for cancer researchers seeking to better model disease and predict patient response to therapeutics 

; however, their use has been limited due to challenging cell culture requirements and a lack of commercially available media systems, according to the company in the release. The off-the-shelf modular tumoroid culture medium kit is designed to fill this perceived gap in the market.

“Cancer’s unpredictable nature presents a significant scientific and clinical challenge for future discoveries and in delivering life-saving treatments to patients,” said Brian Douglass, vice-president and general manager, Cell Biology, Thermo Fisher Scientific, in the release. “Globally, millions of patients and their families still await a cure for cancer despite the decades of advances in research, diagnostics, and treatments aimed at helping lower cancer mortality. We developed the new OncoPro Tumoroid Culture Medium Kit so the scientific community can take greater advantage of tumoroid models to help bring drug candidates to market faster and more cost-effectively.”

“It is well known that tumor organoids are more predictive of clinical outcomes, adverse events and resistance mechanisms than established 2D cell line counterparts,” said Erik Goka, vice-president, Biology, Revere Pharmaceuticals, in the release. “The OncoPro Tumoroid Culture Medium Kit allows us to seamlessly integrate tumoroids into our research and drug discovery processes. With the tumoroid media system, we can propagate all our organoid models in the same culture medium system irrespective of the tissue of origin. This media system has been instrumental in allowing us to bring tumor organoids into our lab for everyday use.”

In addition to the kit, Thermo Fisher Scientific offers services in tumoroid assay development to support outsourced screening and characterization. Customers may also access seven unique OncoProtumoroid cell lines, representing four different cancer indications including colorectal, lung, endometrial, and breast.

Thermo Fisher Scientific’s new tumoroid culture medium kit is designed to help cancer researchers better model the disease.

Thermo Fisher Scientific Releases Tumoroid Culture Medium Kit

Eurofins Genomics Blue Heron, a biotechnology company specializing in advanced gene synthesis, announced the launch of its in-vitro transcription (IVT) messenger RNA (mRNA) synthesis service on June 29, 2023. According to a company press release, the new synthesis service is designed to offer a comprehensive solution for researchers seeking high-quality, customizable mRNA synthesis with fast turnaround times.

According to the release, the new synthesis service is equipped to handle various applications and includes customizable mRNA synthesis capabilities to meet specific research needs. It is designed for use in molecular biology, drug discovery, vaccine development, and gene therapy, among other fields.

News

Eurofins Genomics Blue Heron’s novel mRNA synthesis service is designed for customizable applications.

Eurofins Genomics Blue Heron Launches In-Vitro Transcription mRNA Synthesis Service

Chime Biologics, a China-based contract development and manufacturing organization, announced on June 30, 2023 that it has established three-way strategic collaboration with Nanjing Leads Biolabs (Leads Biolabs), a China-based clinical-stage R&D biotech company, and BeiGene, a biotechnology company with headquarters in China and the United States. The goal of this collaboration is to accelerate the development and global manufacturing of LBL-007, a novel monoclonal antibody (mAb) that was developed independently by Leads Biolabs for cancer treatment.

Phase I clinical trial data of LBL-007 in patients with later-stage solid tumors were announced at the 2021 annual meeting of the American Society of Clinical Oncology, according to a company press release. LBL-007 is currently moving into the clinical Phase II stage.

Leads Biolabs had previously reached an authorization and cooperation agreement with BeiGene under which Leads Biolabs granted an exclusive global development and production license to BeiGene for the mAb, as well as a commercial manufacturing license outside China.

Under the new three-way collaboration, Chime Biologics will cooperate with Leads Biolabs to accelerate the filing of an investigational new drug (IND) application and provide clinical trial materials in China. To achieve IND filing, Leads Biolabs has provided Chime Biologics with its supporting data and information related to the production of LBL-007 and clinical trial materials in mainland China.

Meanwhile, BeiGene and Chime Biologics are also establishing a strategy to facilitate IND applications overseas and clinical trial materials supply outside of China. BeiGene is also providing Chime Biologics support for clinical development and manufacturing of clinical trial materials to enable multiple clinical trials. The companies are seeking indications in colorectal cancer, non-small cell lung cancer, head and neck squamous cell carcinoma, and other solid tumors. According to the press release, clinical trials have been conducted in various countries worldwide, including Australia and the United States.

“We are so pleased to cooperate with BeiGene … and Chime Biologics … LBL-007 is the first innovative antibody drug with global intellectual property which we developed independently. I believe that the development and commercialization of LBL-007 can be expedited through our triangular cooperation, so we can address unmet medical needs and bring hope to cancer patients,” said Kang Xiaoqiang, founder, chairman, and CEO of Leads Biolabs, in the release.

“[LBL-007] has launched nearly ten Phase I and II clinical trials worldwide in the last year, with indications covering colorectal cancer, non-small cell lung cancer, head and neck squamous cell cancer, melanoma, and other solid tumor cancers. I believe that this cooperation will bring new opportunities for the medical community to further conquer cancer,” said Wang Lai, global R&D director of BeiGene, in the release.

Chime Biologics, Leads Biolabs, and BeiGene have formed a three-way collaboration to advance the development and global manufacturing of Leads Biolabs’ lead mAb candidate, LBL-007.

Chime Biologics Forms mAb Development and Manufacturing Deal with Leads Biolabs and BeiGene

SCTbio, a contract development and manufacturing organization (CDMO), and Cyto-care.eu GmbH, a company that specializes in providing advanced solutions for cell therapy and regenerative medicine, have both signed a collaboration agreement to improve the quality of cryopreservation for cell therapy development in Europe. With this collaboration, the companies hope address the need for efficient collection, cryopreservation, quality control, and logistics capabilities of leukapheresis materials to support various aspects of the cell therapy workflow, according to the press release.

A major step for manufacturing advanced cell therapies products (ATMPs) is through the use of cell collection through leukapheresis procedure. Further, the quantity and quality of collected immune cells can directly impact the potency, safety, and efficacy of the final therapeutic product. For this specific collaboration, SCTbio and Cyto-care.eu want to provide a reliable, accessible platform for collecting, characterizing, preserving, and transporting leukapheresis samples.

“We are thrilled to combine both forces and expertise with Cyto-care.eu, a European leader in producing quality blood-based products from leukapheresis, through this strategic partnership,” said Luděk Sojka, CEO of SCTbio, in a press release. “As the cell and gene therapy field continues to advance, the demand for leukapheresis materials is expected to increase. Clinical trials and commercial production of these therapies require a consistent and reliable supply of high-quality blood-derived products to meet patient needs.”

“Through rigorous analysis and in-depth discussions with the experts of the field, the Needle-to-Freezing time has been identified as a crucial factor for success for the development and manufacturing of ATMPs,” said Klaus Fischer, Cyto-care.eu’s managing director, in a press release. “Through this collaboration with SCTbio, we can ensure that our leukapheresis products from patients and from donors continue to meet the high-quality standards of our customers’ requirements.”

The companies hope address the need for efficient collection, cryopreservation, quality control, and logistics capabilities of leukapheresis materials to support various aspects of the cell therapy workflow.

SCTbio and Cyto-care Collaborate to Improve Cell Therapy Cryopreservation 

Verve Therapeutics, a clinical-stage genetic medicines company, announced an exclusive research collaboration with Eli Lilly & Company on June 15, 2023. The deal, which centers around Verve’s preclinical stage, 

 gene editing program targeting lipoprotein(a) (Lp(a)), will see Lilly take over development, manufacturing, and commercialization of Verve’s Lp(a) program following Phase I clinical development. Lilly will pay Verve $60 million upfront, with an additional $465 million plus tiered royalties contingent on various research, development, and commercial milestones.

According to a company press release, Lp(a) is a genetically validated, independent risk factor for atherosclerotic cardiovascular disease (ASCVD), ischemic stroke, thrombosis, and aortic stenosis; this increased risk is most pronounced in individuals with very high Lp(a) concentrations. Verve’s LP(a) gene editing program is evaluated as a treatment for ASCVD.

“Verve was created with a singular focus to protect the world from ASCVD by developing single-course gene editing medicines that address the underlying causal drivers of the disease. Lp(a) is validated as one of these key drivers, and as such, this program represents another important step in our efforts to transform the care of ASCVD,” said Sekar Kathiresan, co-founder and CEO of Verve Therapeutics, in the release. “Blood concentrations of Lp(a) are determined almost entirely by inheritance, and unfortunately, lifestyle and currently approved lipid-lowering therapies have minimal to no impact. In patients with established ASCVD and elevated blood Lp(a), we believe there is a substantial opportunity for a single-course gene editing medicine to permanently lower Lp(a) levels, and we are thrilled to have joined forces with Lilly, an industry leader in cardiometabolic disease, to accelerate this program toward patients.”

“We are pleased to establish this collaboration with Verve, a company uniquely positioned to develop gene editing medicines for common cardiovascular conditions such as elevated Lp(a), a lipoprotein particle that has been established as an important causal and modifiable driver of ASCVD,” said Ruth Gimeno, group vice-president, Diabetes, Obesity, and Cardiometabolic Research, Eli Lilly & Company, in the release. “We look forward to working with Verve to develop a much-needed treatment option for patients with elevated levels of Lp(a).”

Eli Lilly & Company will pay Verve Therapeutics up to $525 million to advance a lipoprotein gene editing research program.

Lilly Enters Gene Editing Collaboration with Verve Therapeutics

Culture Biosciences and Cytiva announced on June 15, 2023 that they are joining forces to advance innovation in upstream bioprocessing through predictive, robust scale-up capabilities, according to a company press release.

“Cytiva is excited to work with Culture Biosciences to help customers gain confidence that the process and data produced from our collaborative platform will help them predict performance at the next level, whether it be clinical or commercial scale,” said Nicolas Pivet, head of automation, controls, digital, and learning at Cytiva, in the press release. “You can accelerate the therapeutic process development.”

Cytiva is focusing on their Bioreactor Scaler tool as a first step to feature Culture’s 250 mL bioreactor. Further, the bioreactor scaler tool helps scaling between development and manufacturing and to de-risk scaling activities and ensure product quality.

“We are thrilled by the prospect to advance upstream bioprocess development with a global leader in the development, manufacturing, and delivery of therapeutics,” said Will Patrick, chief executive officer of Culture Biosciences, in a press release. “Digital technologies have greatly advanced other manufacturing sectors in recent years. We look forward to our work with Cytiva to bring similar innovation to upstream bioprocess development, for the benefit of both our customers and their patients.”

The bioreactor scaler tool helps scaling between development and manufacturing and to de-risk scaling activities and ensure product quality.

Culture Biosciences and Cytiva to Collaborate on Innovative Upstream Bioprocessing 

Charles River Laboratories, a pharmaceutical company specializing in cell and gene therapies, announced the launch of its lentiviral vector (LVV) packaging plasmids on June 13, 2023. According to a company press release, the immediate availability of these off-the-shelf plasmids is intended to provide a streamlined solution for challenges associated with plasmid sourcing for cell and gene therapies, including capacity bottlenecks, timeline pressures, and process development optimization.

“We are thrilled to announce this addition to our product offering,” said Kerstin Dolph, corporate senior vice-president, Biologics Solutions, Charles River, in the release. “Our LVV packaging plasmids allow us to further support advanced therapy programs and most importantly, help expedite the delivery of safe, effective therapies to those who need them most."

“Charles River’s LVV packaging plasmids offer researchers and manufacturers a high-quality, reliable tool to support their LVV viral vectors production,” said Andrew Frazer, associate director, Scientific Solutions, Charles River, in the release. “The new plasmids will be a valuable resource to develop new treatments for a wide range of programs in preclinical and clinical development with the continuity of supply for commercial use”

Charles River’s off-the-shelf lentiviral vector packaging plasmids are intended for use with cell and gene therapies.

BioPharm International's BP Elements, July 2023