Protein therapeutics are an important class of medicines that are used to treat a variety of diseases. According to market research, the protein therapeutics market is estimated to grow at a compound annual growth rate of 6.86% between 2020 and 2027 (1). This growth is expected to be driven by the rapid rise in chronic disorders, advancements in technologies, broadening awareness of protein therapeutics, increasing adoption of plasma-derived therapies to manage chronic disorders, and mounting government initiatives to develop healthcare sectors (1).

However, therapeutic proteins are notoriously less stable than conventional pharmaceuticals; and due to their complex molecular structures, they can be challenging to develop successfully (2). “Compared to small molecules and peptides, proteins are much more complex,” explains Khanh Courtney, PhD, senior director, Biologics, Element. “Therefore, the understanding of the biophysical and biochemical characteristics of a protein biomolecule is critical in the identification of critical quality attributes (CQAs) to ensure consistent safety and efficacy.”

As proteins are formed by long chains of amino acids that interact with each other, secondary and tertiary molecular structures are formed, notes Courtney. Beyond the tertiary structure, the protein can bind to itself, which results in even higher order structures, such as dimer, trimer, and oligomer, she states.

“Certain amino acids could be post-translationally modified with phosphate groups, methyl groups, simple sugars or complex sugars, and so on, leading to a highly complex protein therapeutic molecule,” Courtney says. “The folding of the protein, the higher order structure, [and] the composition of its post-translational modifications, particularly of the glycans, are all examples of protein characterizations that could impact the potency and function of the molecule.”

“Protein characterization is of [the] highest importance throughout all stages of the value chain from discovery to development, through to quality control,” emphasizes Martin Vollmer, strategic program office lead for Biopharma, Agilent Technologies. “Characterization of a protein provides information about its structure and composition, which is directly linked with its function, leading to a greater understanding of differences and ensuring that quality criteria of developed biopharmaceutical products are met according to regulatory guidelines, ensuring drug efficacy and drug safety.”

For Scott J. Berger, PhD, senior manager, Biopharmaceutical Markets, Waters Corporation, whether making a protein drug via a recombinant or synthetic process, development should start with protein characterization and having as much knowledge as possible about the product. “These processes are imperfect, so they are going to make products with low-level variations that are subject to degradation pathways that further raise product variability,” he specifies. “It’s this complexity that creates reliance on sophisticated analytical tools.”

Tools, such as liquid chromatography (LC) and mass spectrometry (MS), have provided process scientists with the ability to monitor important attributes of a molecule in response to changes in the process, Berger adds. “This [capability] gives scientists the flexibility to incrementally refine their process to improve drug quality and yield by establishing analytical comparability,” he says. “Characterization assays often translate to targeted methods for process monitoring and product release where you are verifying that you have control of your process and product quality.”

A key challenge related to protein characterization is that methods usually require a lot of test material, Courtney stresses. “Ideally, protein characterization is done in the early stages of chemistry, manufacturing, and controls (CMC) program development so that critical attributes are identified early on. Unfortunately, the early stage of CMC, for instance during process development, consists of small-scale productions done at the lab bench and have low yield,” she states. “This challenge can be, and is being, dealt with by the advancement of analytical methodologies that can assess more than one characteristic at a time in a single sample preparation or method.”

An example of how this challenge is being overcome can be found in the use of high resolution, high sensitivity MS. “This approach can be used to decipher the peptide map of the protein, post-translational modifications, deamidation, disulfide bonds, aggregation, molecular mass verification, glycan structure elucidation, and even tertiary structure determination, all in one approach,” Courtney explains. “By packaging multiple attributes into one method, a small yield of protein produced at the lab bench could produce a wealth of data to assess protein characteristics to better inform manufacturing and quality decisions.”

There are several immediate challenges to protein characterization in Berger’s opinion. “First, is the increasing complexity of the molecules,” he says. “We’re seeing the appearance of bi- and tri-specific molecules, antibody-drug conjugates [ADCs], and complex fusion proteins, all of which have more attribute variation than your typical monoclonal antibody, requiring enhanced analytics to define and monitor them.”

Additionally, in light of the increasing number of accelerated reviews by regulatory agencies, a challenge has arisen in the form of being able to fully characterize molecules in a compressed timescale and at the same time as stability studies and formulation development are taking place, Berger points out. “So, there is an increasing need for efficient analytical platform methods that can utilize prior knowledge to accelerate these studies,” he states.

“Another challenge many organizations face is the lack of actionable knowledge during the clone selection stage of product discovery/development,” Berger continues. “Use of LC–MS at this stage can ensure that a biosimilar, for example, matches up to the target analytical profile of the innovator molecule when it comes to a set of key and critical quality attributes.” When used as a part of a quality-by-design (QbD) approach, LC–MS can help to ensure that the desired product attributes are built into the molecule, he notes. If there is insufficient knowledge about a molecule at this stage of development, it is possible that impurities or instabilities won’t be discovered until after the product is placed into clinical production, Berger warns.

For bench scientists, experimental bias can be a significant challenge, Berger highlights. “A top reason why assay-to-assay test results vary from one scientist to another has to do with complicated and multi-step sample preparation methods,” he says. “For multi-attribute monitoring of biomolecules by LC–MS, it’s important that you have a very high-quality and reproducible sample for meaningful results. To address this need automated sample prep technologies are being investigated by many groups as a tool to improve sample quality and reproducibility by eliminating the human factor from potentially biasing assay results.”

Undesirable secondary interactions can adversely impact analytical data when analyzing organic acids, organophosphates, oligonucleotides, phosphopeptides, acidic glycans, and phospholipids, Berger notes. “One example of a secondary interaction is the non-specific adsorption by certain biomolecules to the metal surfaces—or even to bio-compatible materials like titanium—of liquid chromatographs and chromatographic columns,” he states. “Another type of secondary interaction is caused by the highly active surfaces of proteins which have a propensity to interact with the hydrophobic and electrostatically-active sites on the silica and hybrid silica chromatographic particles in the UHPLC [ultra-high performance LC] or UPLC [Ultra-Performance LC] column.”

These secondary interactions cause difficulties when characterizing and monitoring drug products by size exclusion chromatography (SEC), which is used to determine protein sample size variants claims Bill Warren, Principal Bioseparation product manager, Waters Corporation. If these attributes are not carefully controlled then the safety and efficacy of the product can be adversely affected, Warren specifies. “It’s for this reason that regulatory agencies require drug firms to accurately quantify the protein size variants like mAb aggregates, monomers and fragments in protein drug products,” he says.

Vollmer emphasizes that protein characterization is highly complex and is only becoming more complex over time. “In the past, it was mainly monoclonal antibodies (mAbs), hormones, vaccines, modified human proteins, and similar therapeutic proteins. Now with the discovery of antibody [drug conjugates (ADCs) fusion-proteins, bi-specific Abs, and other modalities, the task of protein characterization now requires a much more flexible approach,” he asserts.

This increasing complexity is then also tied to the skill of the analytical scientist that needs to perform the characterization, Vollmer stresses. “There are a multitude of assays that need to be performed and the instrumentation required is more sophisticated, but costly,” he explains. “Big hurdles are, therefore, the cost of implementation, operation, and skilled personnel.”

“Another important challenge is the large amount of data that needs to be analyzed for meaningful information and further action,” Vollmer continues. “More broadly, regulatory hurdles and inertia in the industry are often roadblocks for fast innovation.”

“LC and MS have been the empowering technologies for the well-characterized biotherapeutic,” Warren says. “LC is essential for fully analyzing an intact protein or its sub-units for glycan profiling or peptide mapping or for looking at protein charge or size variants.”

A technique that is useful for characterizing protein size variants is SEC, Warren adds. There have been several new SEC columns that have been brought to the market recently, he states, that address “the problem of secondary interactions (both ionic and hydrophobic) when separating protein aggregates, monomers, and fragments over a molecular weight range of 10,000 to 6,500,000 Daltons.”

Berger also notes that through combining LC and MS, it is possible to establish the primary structure (product sequence), which is a regulatory filing requirement. “With LC–MS, you get to explore the product variation, its stability, and degradation pathways, which ‘hot spots’ to monitor to ensure that the product qualities and your processes are under your control,” he says. “LC–MS is also ideal for looking at a molecule’s higher order structure, which relates to the folding and stability of these biotherapeutics, and how they interact with their protein targets, or themselves when it comes to things like molecular aggregation.”

The structural information obtained by LC–MS can drive product development and can also form the basis of intellectual property, Berger points out. “Understanding higher order structure and stability dynamics may require the use of techniques like hydrogen deuterium exchange or ion mobility mass spectrometry (IMS-MS) and more recent techniques, such as collision-induced unfolding (CIU). CIU is an approach where the mass spectrometer is almost like a calorimeter, adding energy to the protein molecule and seeing how it unfolds within the IMS-MS,” he says. “These techniques allow you to ask questions that go beyond primary structure to understanding the dynamics of its folded structure and interactions with its protein target.”

Courtney highlights three techniques that are important for protein characterization: “High resolution, high sensitivity, orbitrap MS for protein and glycan identification, and structural elucidations; capillary electrophoresis to determine charge heterogeneity; and circular dichroism for secondary structures.”

Although there is no single standard technique to characterize proteins, Vollmer concurs that LC and high-resolution MS are regarded as the gold-standard thanks to the “rich” information such techniques offer on physiochemical properties of the protein. “Other alternative techniques are capillary electrophoresis (CE) and CE–MS because they can provide excellent resolution,” he adds.

“Spectroscopy, such as ultraviolet, Raman, near infrared, or fluorescence, are also widely applied to determine more specific requirements, such as concentration or purity. These techniques are fast and can even be applied inline,” Vollmer concludes. “Cell analysis technologies, such as metabolic analyzers, real-time cell analyzers, cell-imagers, and cell counters are applied to characterize suitable clones and host cells for protein manufacturing.”

Protein Therapeutics Market Research Report: by Type, Application, Protein Function, End User—Forecast to 2027

, Market Report (February 2021).
2. A. Bolje and S. Gobec, 

Felicity Thomas is the European editor for the Pharmaceutical Technology Group.

Articles

Features

The challenges of conducting protein characterization can be overcome through the use of technologies that empower scientists in their analytical work. 

Protein Characterization Strengthened by Enabling Technologies

Chimeric antigen receptor (CAR)-T cells have already changed how certain cancers are treated. As of February 2022, FDA had approved five CAR-T cell therapies to treat liquid tumors in patients with various types of leukemia and lymphoma (1). The biopharmaceutical industry has invested heavily in the future of this novel treatment modality, and CAR-T cells now account for 56% of cell immunotherapies in development (2). By 2030, one forecast predicts that the CAR-T cell market will grow to $21.7 billion (3).

With this growth, however, comes the realization across the industry that CAR-T cell therapy development is a complex process that requires standardization, regulation, and meticulous quality control to reliably generate safe and effective therapies. In addition, when developing CAR-T cells, biomanufacturers can have their choice of cell sources and bioprocesses, which can lead to variability in success as well as openings for inefficiency and waste. Analytical tools such as droplet digital polymerase chain reaction (ddPCR), a technology platform used to quantify nucleic acids, can support CAR-T cell manufacturers in developing successful CAR-T cell products.

The importance of quality control in CAR-T cell development is evident in the development process itself. First, a physician draws blood from a patient and a technician extracts the T cells. Next, a technician delivers the 

 gene to the T cells, typically using viral vectors. The cells take up the gene and express it, generating the CAR protein. This protein, in turn, gets shuttled to the cell surface,` thereby transforming the T cell into a CAR-T cell. Finally, the technician amplifies the CAR-T cells in a bioreactor and delivers the batch to the clinic, where a physician transfuses the cells back into the patient.

Each step in this process depends on nature: technicians rely on viruses and T cell machinery to generate CAR-T cells as well as on the proper environmental conditions to expand the cells, while physicians rely on the patients’ bodies to tolerate the cells and degrade them at an appropriate rate. Because developers and physicians cannot fully control all of these factors, they must monitor the creation and administration of CAR-T cells to ensure they perform as expected and will not induce adverse reactions in patients. Below are several sources of variability in CAR-T development and how ddPCR technology can help resolve some of the uncertainty inherent in the process.

In the development of CAR-T cells, one key initial step is using viruses, such as adeno-associated virus (AAV) or lentivirus, 

 gene. But once the initial conditions are set, it is difficult to control the behavior of these viruses and the genetic sequences they contain. As a result, the number of 

 transgenes that enter each cell and integrate into the genome can vary. Depending on the success of this transfection step, the treatment may not work at all, or worse, it may elicit a systemic inflammatory response that can cause further harm to patients (4). To protect patients, FDA recommends that CAR-T cell manufacturers performing viral transduction screen out cells containing less than one or more than four transgene copies per cell (5). To do this screening process, technicians need to quantify the 

 transgene copy number accurately and precisely.

Quantitative polymerase chain reaction (qPCR) is the standard method for quantifying nucleic acids, but it is not sensitive enough to quantify 


transgenes with the accuracy and precision needed to satisfy regulators. The qPCR technique quantifies nucleic acids indirectly, using a standard curve to estimate copy number. Scientists must generate this standard curve using serial dilutions, a process that is time-consuming and prone to human error. This error creates variability that makes it challenging to quantify small copy numbers. If developers cannot tell with certainty whether or not a CAR-T cell batch contains at least one transgene copy per cell, they cannot be sure whether their transfection method was successful.

ddPCR technology, however, can offer precise measurements of transgene copy number because of its ability to quantify gene sequences directly without using a standard curve. The technique involves partitioning a sample into approximately 20,000 nanoliter-sized droplets that contain one or a few nucleic acid strands each, with some droplets containing the target gene and some not. A separate PCR reaction takes place in each one. Starting with a sample of nucleic acids from CAR-T cells and using primers targeted to the 

 transgene, DNA amplification will only occur in the droplets containing that gene. As the gene amplifies, a reporter gets cleaved from the probe and emits a fluorescent signal. The fluorescent droplets can be counted to derive the transgene concentration in the original sample. Unlike qPCR, ddPCR does not measure the degree of amplification to estimate copy number in the original sample; instead, it counts whether or not amplification took place, providing a digital measure of copy number. This ability makes ddPCR technology more accurate and precise and, therefore, more reliable for assessing CAR copy number in CAR-T cells.

These attributes were quantified in a study performed by 

 at the Huazhong University of Science and Technology in Wuhan, China (6). The researchers directly compared qPCR and ddPCR instruments in their ability to quantify DNA standards and CAR transgene copies in clinical peripheral blood samples from patients on CAR-T therapy. They found that ddPCR demonstrated greater sensitivity than qPCR. Specifically, among a series of DNA standards, ddPCR assays detected as few as 3.2 copies/mL, while qPCR showed a negative result at that concentration. Furthermore, their ddPCR assay could reliably detect as few as five copies per reaction among clinical blood samples, while qPCR could not detect concentrations lower than 20 copies per reaction. Additionally, ddPCR technology showed lower intra-assay and inter-assay coefficients of variance for the series of diluted standards than qPCR, suggesting that ddPCR is more repeatable and reproducible.

Meanwhile, researchers at the National Institutes of Health Clinical Center wanted to see if this method of copy number quantification was robust. To do this, 

 tested the consistency of ddPCR assay results following several changes to the development workflow (7). Specifically, the researchers transfected T cells using either lentiviral or retroviral vectors. They then assessed transfection success right away or after the cells had been frozen for three or six weeks, using three different technicians across two different laboratories. When they quantified 

 copy number amidst all of these protocol variations, their ddPCR assay delivered consistent results.

ddPCR technology was also central to optimizing 

’s transduction protocol. The researchers varied the viral vector-to-T cell ratio as well as varied their centrifugation protocol (i.e., either centrifuging their samples or not) following transduction to identify the best protocol for maximizing transduction efficiency and copy number per cell. They used flow cytometry to measure the effect of these variables on transfection efficiency while using ddPCR technology to measure copy number. Both of these measures were correlated throughout the experiments. Transfection efficiency and copy number per cell increased as the vector-to-target cell ratio increased, but only when the samples were not centrifuged. Conversely, centrifuging the samples increased transfection efficiency at lower vector-to-target ratios. Overall, these data demonstrated that ddPCR technology can reliably quantify 

 transgene copy numbers across different conditions and can be used to assess the impact of various protocol deviations on the success of CAR-T cell production.

 gene can lead to immune reactions, but this scenario is not the only case in which a CAR-T cell could theoretically pose a risk to patients. For example, CAR-T cell production using lentivirus could potentially create replication-competent lentiviruses (RCL) that could infect patients and induce the rapid growth of T-cell neoplasms. This scenario is purely theoretical; however, to be safe, FDA recommends that manufacturers test their clinical vector lots and manufactured cell products for RCL. In addition, the agency recommends that doctors test their patients’ blood after receiving CAR-T cell therapy (8).

ddPCR technology can be used to detect RCL. In one study, researchers 

 at the Mayo Clinic in Rochester, Minn., developed a ddPCR assay that detects the envelope sequence of the virus (9). The limit of detection for their assay was 10 copies/μL. The technique, overall, provided sensitive, specific, and reproducible RCL detection, expediting quality-control assessments compared to traditional cell-based or qPCR-based approaches. 

’s results suggest that ddPCR could be used to screen out CAR-T cells that contain RCL so that these cells never reach patients.

Physicians infuse patients with a specific dose of CAR-T cells that is intended to deliver a therapeutic benefit without causing harm to patients. However, there is no way to fully predict CAR-T cell kinetics once the cells enter the body. Consequently, physicians must monitor CAR-T cell concentration in the blood on a serial basis to ensure the cells are present in the appropriate concentration for the right amount of time.

CAR-T cells are supposed to survive in the body for a few months, but, if they persist for too long, they can elicit a prolonged inflammatory response even after they’ve effectively treated the patient’s cancer (4). Conversely, if the cells degrade too quickly, they will not do their job. Therefore, physicians must monitor CAR-T cell kinetics to optimize clinical follow-up regimens, understand treatment failure, and understand the cause of adverse events.

There are currently no standard methods for measuring CAR-T cell kinetics, but ddPCR assays present an attractive option. Because ddPCR technology can detect small concentrations of nucleic acid sequences in liquid samples, it can be used to precisely quantify CAR-T cell concentration in the blood. Furthermore, because liquid biopsies based on ddPCR technology are minimally invasive, these tests can be run on a serial basis without causing further harm to patients.

 in Germany demonstrated the feasibility of using ddPCR technology for this purpose in patients with various types of lymphoma (10). Two CAR-T therapies, tisagenlecleucel and axicabtagene ciloleucel (axi-cel), are currently approved to treat this disease. In patients with lymphoma treated with CAR-T cell therapy, plasma CAR-T cell concentration correlates with treatment response and patient outcomes, indicating that CAR-T cell kinetics can serve as a predictive biomarker of therapeutic efficacy (11–13). To test this hypothesis, researchers developed a ddPCR assay that examines axi-cell kinetics. First, they tested it on positive control samples containing various known concentrations of CAR-T genomes. Their assay reliably detected one CAR gene per genome.

The researchers then tested 54 samples from seven patients between four weeks and nine months following treatment. They found that CAR-T cell kinetics varied greatly between patients, demonstrating the importance of serial monitoring. Looking closer at each patient, the researchers discovered that CAR-T cell persistence and peak expansion correlated with clinical responses. For example, the patient whose 

 levels dropped to undetectable levels at day 75 after treatment showed no clinical improvement, while the patients whose 

 levels were still detectable after nine months went into remission. At the other end of the spectrum, the patient with the highest peak 

 copy levels experienced severe side effects following therapy. Altogether, these data revealed the variability in CAR-T cell kinetics and the importance of serial monitoring.

ddPCR and CAR-T cell therapy go hand-in-hand

ddPCR technology has already proven useful in multiple research areas over the past decade. CAR-T cell therapy is a key current example that demonstrates the technology’s full potential to help biopharmaceutical manufacturers and physicians keep patients safe and healthy. CAR-T cell therapy development is not simple, but by quantifying 

 transgenes and other genetic elements using ddPCR assays throughout the process, CAR-T developers will be more successful in developing therapies that will achieve regulatory approval, and physicians can be more confident that they can make a meaningful difference in their patients’ lives.

1. University of Pittsburgh Medical Center Hillman Cancer Center, “FDA-approved CAR T-Cell Therapies,” 

2. Cancer Research Institute, “Cancer Cell Therapy Landscape,” 

CAR T-cell Therapy Market - Global Industry Analysis, Size, Share, Growth, Trends, and Forecast, 2021-2031

8. US Department of Health and Human Services, FDA, CBER, 

Mark White, PhD, is the associate director of Biopharma Product Marketing, and Marwan Alsarraj is the Biopharma Segment manager; both are at Bio-Rad.

Standardization is needed in CAR-T cell therapy development due to its inherent complexity.

Using ddPCR to Asses CAR-T Cell Quality

Research from the University of Michigan has demonstrated that twirling infrared light can be used to probe the structures of molecular crystals. The finding, reported in a March 21, 2022 announcement issued by the university, addresses one of the difficulties associated with drug development, which is determining if drugs and supplements with chiral structures are turning in the correct direction.

Drugs containing molecules with the wrong twist can cause unpleasant or dangerous side effects. According to the university’s announcement, this method uses terahertz radiation, a portion of the infrared part of the spectrum, to quickly recognize wrong twists and wrong chemical structures in packaged drugs.

Researchers hope that the technique can also be used to find harmful accumulations of twisted molecules in the body. This could help diagnose bladder stones, insulin fibrils, and amyloid aggregations such as the plaques that appear in Alzheimer’s disease, according to the university’s announcement.

“We foresee new roads ahead—for instance, using terahertz waves with tailored polarization to manipulate large molecular assemblies,” said André Farias de Moura, professor of chemistry at the Federal University of São Carlos, in the announcement. “It might replace microwaves in many synthesis applications in which the handedness of the molecules matters.”

The technique was developed by an international team consisting of researchers from Federal University, the Brazilian Biorenewables National Laboratory, the University of Notre Dame, and Michigan State University.

New Technology

Research from the University of Michigan showed that Terahertz light can be used to probe both the structures of molecular crystals and their twists.

Twisted Vibrations Enable Quality Control for Chiral Drugs and Supplements

Thermo Fisher Scientific introduced a new large-volume electroporation system on March 16, 2022 that is intended to enable the development of cell therapies. The system’s goal is to allow cell therapy developers to move more easily from clinical development to commercial manufacturing.

The Gibco CTS Xenon Electroporation System features a closed, highly flexible design to facilitate gene modifications without the use of traditional viral vectors. According to a company press release, the electroporation approach enables the introduction of a payload into a cell by temporarily increasing cell permeability; this is done using an electrical pulse.

Unlike existing large-volume cell therapy electroporation solutions on the market, Thermo Fisher’s new system features programmable, flexible electroporation conditions and offers cell therapy developers full control to optimize a variety of hard-to-transfect cell types and payloads. The system can be used as a standalone technology, or it can be integrated with the Gibco CTS Rotea Counterflow Centrifugation System as part of a modular, closed, and automated cell therapy manufacturing workflow, according to Thermo Fisher.

The CTS Xenon Electroporation System can transfect up to 2.5e9 cells in 25 mL in a sterile system. This transfection capacity enables rapid and efficient non-viral transfection for clinical manufacturing applications. In addition, the system delivers up to 80% cell viability and 90% gene knockout transfection, allowing scale up from the small-volume Neon Transfection System without the need to re-optimize the electroporation parameters. Thus, the system is designed to easily transition from benchtop to clinic, according to the company.

“We launched the CTS Rotea System in 2020 as part of our strategy to deliver fit-for-purpose cell therapy solutions that can easily scale from development to commercialization. Adding the CTS Xenon Electroporation System to our end-to-end workflow will help customers overcome manufacturing challenges that hinder production, enabling them to bring life-saving therapies to patients faster,” said Amy Butler, president of biosciences, Thermo Fisher Scientific, in a company press release.

Thermo Fisher Scientific’s new Gibco CTS Xenon Electroporation System aims to provide easier scale up for cell therapies, from clinical development to commercial manufacturing.

Thermo Fisher Introduces New Electroporation System for Large-Scale Cell Therapy Development and Manufacturing

ATCC, a global biological materials management and standards organization, announced on March 28, 2022 that it has been awarded a two-year task order by the National Cancer Institute (NCI). Valued at $5.6 million, the task order is part of an existing indefinite delivery/indefinite quantity (IDIQ) contract that ATCC has with NCI. The IDIQ contract is for supporting the Division of Cancer Epidemiology and Genetics (DCEG) Molecular Epidemiology Assay Support (MEAS) program.

Under the contract, ATCC may coordinate receipt of specimens from clinical and field sites as well as process fresh samples within 24 hours. ATCC will also perform a broad range of molecular assays to characterize the models as required. ATCC may also provide support services for the procurement, storage, and shipment of specimens to and from international and domestic specimen sites as well as prepare specimens for transport to testing laboratories by aliquoting, labeling, and batching, as needed, according to a company press release.

DCEG’s mission is to discover the causes of cancer and research ways to prevent cancer by conducting transdisciplinary epidemiological and genetic research. DCEG investigators typically rely on contract services to provide technical support for specimen collection, receipt and processing, and assistance in identifying laboratories to perform state-of-the-art testing using high-quality biological specimens collected from clinical and field sites. The MEAS program provides support services for clinical studies on the genetic and other cellular events that influence the onset of different types of cancer.

"At ATCC, we are committed to supporting members of the clinical research community by providing them with the highest quality biomaterials for cancer research," said Raymond H. Cypess, chairman and CEO of ATCC, in the press release. "We know how important this is when they are designing studies to discover cancer's genetic and environmental determinants and innovative approaches to cancer prevention. Scientific research is critical for unlocking incredible breakthroughs in global health."

"Through our expertise in providing standard reagents, functional testing, and biorepository operations and management, ATCC continues to support cancer research through DCEG's clinical studies," said Joseph Leonelli, senior vice-president of ATCC Federal Solutions, in the press release. "We look forward to our continued relationship with NCI through the MEAS program."

News

Under the two-year task order, ATCC will continue to support the DCEG MEAS program in receiving, processing, storing, analyzing, and distributing clinical specimens from different cancer types.

NCI Awards ATCC Task Order to Provide Cancer Epidemiology Services to Research Community

Spectrum Chemical, a supplier of fine chemicals and laboratory equipment, announced twelve new products in its bioCERTIFIED product line on March 15, 2022. Launched in 2019, bioCERTIFIED is a quality management system and chemicals portfolio consisting of products designed to reduce time to market for new drugs and therapeutics while still meeting regulatory and compliance requirements.

The new products include sugars/carbohydrates, organic and inorganic salts, and preservatives used in various bioprocessing applications—including upstream cell culture development, downstream purification, polishing, and final fill production. The new product list includes Sodium Chloride United States Pharmacopeia–European Pharmacopeia (USP-EP), an inorganic salt with multiple applications ranging from cell culture to downstream purification and polishing, and Sucrose (beet-derived) USP-EP, National Formulary, Japanese Pharmacopeia, and Chinese Pharmacopeia, a sugar/carbohydrate for upstream cell culture and downstream final fill.

“We have a 50-year track record providing products with comprehensive scientific documentation to quality-focused industries such as pharma and biopharmaceuticals,” said Russell Kneipp, chief operating officer, Spectrum Chemical, in a company press release. “All bioCERTIFIED products carry our ‘100% tested, 100% transparent, 100% trusted’ guarantee, reflecting our ongoing commitment to support biopharmaceutical development and production of biologics for new vaccines, therapeutics, and an array of future applications.”

“Spectrum Chemical has decades of experience dealing with the ever-changing global regulatory environment,” said Jigisha Patel, vice-president of Global Regulatory and Compliance, Spectrum Chemical, in the press release. “Due to COVID-19, there is an even greater worldwide demand for chemicals whose quality and safety are thoroughly documented. Not only do our chemicals meet regulatory and compliance requirements, our documented change controls and sourcing protocols ensure full supply chain transparency.”

Spectrum Chemical is adding twelve products to its bioCERTIFIED product line.

Spectrum Chemical Launches New bioCERTIFIED Products

ProteoGenix, a France-based biotech contract research organization, announced the launch of XtenCHO Transient Expression System on March 15, 2022. This new expression system can achieve up to ten times higher yields with less hands-on time when compared to existing solutions because of enhanced plasmid stability and optimized metabolism, according to a company press release.

ProteoGenix developed XtenCHO from the parental cell line Chinese hamster ovary (CHO)-K1 by engineering its genome to exhibit properties that maximize protein production yields. An animal component-free growth medium, highly efficient transfection reagent, and ultra-high expression vector work together to reduce premature plasmid loss, creating a three to ten-fold increase in transient gene expression yields compared to market-leading CHO systems. The company also states that the product has an enhanced robustness that allows it to recover quickly after thawing and perform satisfactorily in suboptimal conditions.

“The new XtenCHO system produces monoclonal antibodies that are indistinguishable from biopharmaceutical-grade biologics produced in stable systems,” said Raphaël Hopfner, chief scientific officer and co-founder, ProteoGenix, in the press release. “This eases process scalability and reduces the risks when transitioning from the bench to the clinic.”

“We have launched XtenCHO to help speed up the process of early drug development; making it safer, more flexible, accessible and easier to integrate into existing biologics discovery workflows using cost-effective reagents to achieve very high yields,” said Philippe Funfrock, CEO and co-founder, ProteoGenix, in the press release.

ProteoGenix’s new host has been used and optimized by ProteoGenix for approximately five years, creating more than 3500 recombinant proteins. This includes antibodies from multiple species, isotopes, and formats, including bispecific antibodies, antibody drug conjugates, and nanobodies. Non-antibody proteins, such as highly functional enzymes, membrane-bound receptors, cytokines, interleukins, and transcription factors, have also been produced.

ProteoGenix’s new XtenCHO Transient Expression can achieve up to ten times higher yields than market-leading CHO systems.

ProteoGenix Launches New Transient Expression System

After months of negotiations, Congress approved a massive federal government spending package in March 2022, spurred by bipartisan support for including $13.6 billion in aid for Ukraine. The $1.5 trillion omnibus funding bill for fiscal year 2022, which began Oct. 1, 2021, increases funds to combat the opioid crisis and extends telehealth provisions, while also directing resources to a broad range of programs to advance health and medicine.

The measure increases FDA’s budget by $102 million to $3.3 billion in discretionary budget authority. FDA’s total funding amounts to $6.1 billion, with almost half of that—$2.8 billion—provided by approved user fees. The boost in federal funds is smaller than proposed earlier by Congressional committees, but sufficient to “reflect continued Congressional support for a strong, well-funded FDA,” commented 

The added $102 million allots $29 million to medical product oversight, $29.5 million to food safety activities, and retains $41 million for cross-cutting initiatives. The legislators direct FDA to use the increase for medical products to better track medical device shortages, tackle the opioid crisis, provide unannounced inspections of foreign drug manufacturers, bolster orphan and rare disease drug development, and support FDA data modernization, field inspections, and minority health programs. The funding committees highlight the importance of conducting real-time, unannounced plant inspections and instruct FDA to spend $5 million on hiring additional foreign-based investigators for this purpose.

are provided in the report from the House Appropriations Agriculture subcommittee. Of the $2.8 billion to be collected in user fees, $1.2 billion comes from fees on drugs and biologics, $540 million from generic drugs and $40 million from biosimilars. The budget plan calls for FDA to do more to regulate cannabis-derived products, advance strategies to combat the opioid crisis, develop a comprehensive food traceability system, and take steps to protect against adulterated dietary supplements.

In the broader health and medicine arena, 

 increases support for the National Institutes of Health (NIH), the Centers for Disease Control and Prevention (CDC) and other agencies in the Department of Health and Human Services (HHS). Of note, Congress provides $1 billion to establish the Advanced Research Projects Agency for Health (ARPA-H) as part of NIH. The amount is way less than the $6.5 billion initially sought by the Biden administration for this new entity, but an important first step in moving it forward.

More controversial is the Democrats’ decision to approve the broad spending package without including President Biden’s $30 billion request for additional COVID-19 relief funds. That amount had already been reduced to $15.6 billion to purchase and continue research on more anti-COVID vaccines, therapies, and test kits, but the provision was dropped due to dissension among Democrats over the proposed cuts. Budget increases at CDC, NIH, and the Biomedical Advanced Research and Development Authority (BARDA) are expected to support further R&D in this area, pending future agreement on COVID research activities, but researchers fear significant reductions in these programs.

At the same time, the broader budget bill provides support for a host of specific initiatives, such as a measure limiting access to flavored vaping products by clarifying FDA authority to regulate these products. There are provisions to prevent domestic violence and billions to support thousands of pet projects favored from individual legislators.

Resolution of this federal spending bill clears the way for Congress to start considering legislation to reauthorize a packet of FDA user fee programs by Sept. 30, 2022. That deadline opens the door for the legislators to include multiple proposals affecting FDA and biomedical research in the fee package, as seen in a long list of bills already being considered by the House Energy & Commerce Health subcommittee.

The proposals include the Cures 2.0 Act (HR 6000) to advance a range of biomedical research programs plus multiple measures that fall under the heading of “encouraging innovation and improving oversight.” There are bills to 

overhaul FDA’s accelerated approval process

, encourage drug production in the United States, advance diversity in clinical trials, support development of certain orphan drugs and rare disease treatments, and ensure access to affordable medicines. The House panel will begin the evaluation process for these and other proposals at a hearing March 17, 2022.

The US Congress approved a federal spending package that increases funding for a range of programs to advance health and medicine.

Federal Budget Bill Boosts FDA Funding, Supports New Research Institute

Flow Eighteen38, a protein sciences company, announced a €5 Million (US$5.58 million)investment into its protein production and biophysical characterization capabilities on March 8, 2022. The investment comes from its parent company, FairJourney Biologics, a company specializing in the discovery and optimization of antibodies.

According to a company press release, the investment will be used to develop a new laboratory that will be designed to double the company's protein purification capacity. These facilities will accommodate 15+ high-end fast protein liquid chromatography systems, all of which will be individually optimized to scale-up custom production, support unique partner requirements, and fast-track standard productions.

The investment will also be used to purchase:

LabChip’s GX Touch for capillary electrophoresis sodium dodecyl sulfate (CE-SDS)

Thermo Scientific’s Vanquish ultra high-performance liquid chromatography (UHPLC)

Endosafe nexgen-MCS for endotoxin testing

Two BioAccord liquid chromatography–mass spectrometry (LC–MS) systems.

The funding will also drive further recruitment for production and characterization services and cold storage and logistics solutions. It will also contribute to continued supply of quality antibody reagents that comply with new European Union recommendations.

“This investment is further commitment to our partners to ensure we remain on the cutting-edge of protein production and biophysical solutions and to continue to deliver quality results our diverse portfolio of partners have come to know and expect,” said António Parada, CEO, FairJourney Biologics, in the press release.

“We are thrilled for this new stage of Flow Eighteen38,” said Teresa Barata, head of Protein Science Division, Flow Eighteen38, in the press release. “The new investment will enable us to continue to support our clients at the various stages of the discovery process and extend it to pre-[chemistry, manufacturing, and controls] stages, for faster and successful biotherapeutics development.”

Flow Eighteeen38 is receiving a €5 million (US$5.58 million) investment from its parent company, FairJourney Biologics.

Pharmaceutical Technology's In the Lab eNewsletter, April 2022