Determining product quality is a critical measure of a proposed biosimilar product. Appropriate analytical methods are needed to assess quality as well as for applying quality control (QC). One of the key challenges in developing or selecting an assay to determine biosimilar product quality is determining which critical quality attributes (CQAs) are the most important for a given stage of the drug development lifecycle, notes Stephen Shakespeare, product manager, Sartorius.

“For example, a binding assay may be more relevant during clone selection than a mechanism-of-action-bioassay, which would be more suited for the final stages of development,” Shakespeare says.

Titer is another critical process parameter, and to ensure final product quality, it is necessary to measure the product concentration throughout a bioreactor run, adds Laura Madia, sales and marketing manager at IDEX Health & Science.

Another point of consideration is the kind of information that is required from the assay. “Is it selective and specific for the product? Does it address the CQAs that have been identified from analysis of the innovator?” says Shakespeare, who points out that the use of orthogonal methods provides a host of information on the product that is critical for ensuring the quality.

“There are a number of challenges facing biopharma production, and their impact (overall production, development timelines, cost, etc.) can be more significant for companies developing biosimilars,” includes Madia. “One challenge in particular is the drive for improving process efficiencies. Biosimilars are considered to be cost-effective alternatives to expensive biologics. There is a significant pressure placed on reducing production costs through improved manufacturing processes to improve yields without introducing product variation that might require additional processing or testing.”

The tradeoffs between time and cost are particularly important when developing biosimilars given the limited development time, Madia continues, saying that “biosimilars do not benefit from the same patent protection as novel biologics so there is typically more pressure on prices immediately after approval.”

Another question to consider is what are the most effective assays currently used to test for biosimilar product quality? When assessing a biosimilar, Shakespeare explains, it is critical to take what he calls an orthogonal approach to looking at antibody fragment (Fab) binding and fragment crystallization (Fc) binding using a variety of techniques.

“In Fc functional assays, for example, antibody-dependent cellular cytotoxicity (ADCC) can be addressed in numerous ways using peripheral blood mononuclear cells (PBMCs) or natural killer (NK) cells isolated from blood. Meanwhile, reporter gene assays can be validated for submission to the regulators. The sensitivity of each of these methods provides key information on the quality of the product at different stages of development,” Shakespeare notes.

He adds that there is an array of methods, from enzyme-linked immunosorbent assay (ELISA) and flow cytometry to surface plasmon resonance (SPR), that can be used to determine Fab binding.

Another example of an orthogonal approach is the use of size exclusion and ion exchange analytical methods, which can show differences in aggregation/fragmentation and charge profile, respectively. These attributes can be important when assessing the stability and safety of a biosimilar, Shakespeare says.

Understanding the innovator biologic is also critical to understanding the CQAs of the biosimilar product, Shakespeare also notes. Understanding the innovator CQAs allows the biosimilar developer to determine the quality target product profile (QTPP) design space. Design-of-experiment software (e.g., Umetrics software, Sartorius) enables biomanufacturers to monitor the production process and track whether certain biosimilar CQAs are drifting, allowing them to respond accordingly.

“Being able to monitor different aspects of a biosimilar production in real-time, such as binding and function, enables you to control CQAs and minimize any potential downstream issues with delivering consistent quality product,” Shakespeare states.

There are situations where a process depends on a quick measurement to move forward, Madia remarks. “Measuring titer is not considered simple nor quick. Titer is often run on a high-performance liquid chromatography (HPLC) in the QC lab due to the high level of skill required for method development and day-to-day operation. It is not uncommon for upstream process development or manufacturing teams to wait 24 hours or longer to see the results of a HPLC titer analysis because they had to send it to a separate lab,” Madia explains. As a result, titer is often underutilized or batched for retrospective insights, rather than using titer for real-time decision making, Madia points out.

Since the manufacture of the first biosimilars, there have been many innovations in analytical techniques and practices. In recent years, for example, high throughput (HTP) flow cytometers (e.g., iQue screener, Sartorius) have enhanced the speed with which data can be generated, Shakespeare highlights. HTP flow cytometry, coupled with live cell imaging (e.g., Incucyte, Sartorius), enhances the breadth of information that is collected and allows for more informed decisions to be made faster, he explains.

Other recent analytical innovations include capillary electrophoresis techniques, which have replaced slab gel techniques, resulting in assays that have high resolution, are highly reproducible, and are applicable to a wide range of protein molecules—not just immunoglobulins and biosimilars. “The use of capillary isoelectric focusing (cIEF) and capillary electrophoresis sodium dodecyl sulfate (CE–SDS) techniques are now almost universally applied for CQA determination, stability studies, and lot release testing,” Shakespeare says.

Manufacturers, meanwhile, are continuing to implement liquid chromatography–mass spectrometry (LC–MS) techniques for investigating glycan structure, verifying sequences, analyzing intact mass, and analyzing post-translational modifications of the innovator and of biosimilar candidates, Shakespeare adds. “By introducing simple and affordable kits, manufacturers are getting greater instrument resolution, improvements in software processing and databases, and fast sample preparations,” he explains.

“In a similar vein, the ongoing reduction in particle size to < 3 µm in ultra-high-performance liquid chromatography (UHPLC) columns continues to enable reduced run times and higher throughput while maintaining peak resolution,” Shakespeare also adds.

Having an innovative new product that measures titer on demand is also a beneficial advancement, Madia adds. Drawing on experiences with the Tridex Analyzer (IDEX), Madia highlights how innovative technology can enable development and facilitate experimentation by providing trend data quickly and easily. With the trend data, it may be possible to determine product loss during the evaluation of filtration options or to monitor breakthrough with a new piece of equipment. “Ultimately, a full analysis will be required as part of the product acceptance, but by eliminating options early, it is only necessary to run a full analytical evaluation on the strongest candidates,” Madia says.

The most common application for antibody titer is in determining concentration in the bioreactor tank at the end of a cell culture run, Madia emphasizes. Once a process moves to production, current analytical tools are too large or complex to be co-located near the tank or in the manufacturing suite, she explains.

“During the crucial transition from upstream to downstream purification, manufacturing teams require the final titer data in order to load the purification columns. Once the tank is emptied and the filtration step is complete, production stops. Product is put in a holding tank while the team waits for results. Eliminating downtime while waiting for titer could shave hours or days and can also reduce the risk of moving product between areas. Using real-time titer to streamline this transition can make a real impact at manufacturing scale,” Madia says.

In May 2019, FDA proposed a draft guidance for the development of biosimilars (1) in which it outlines comparative analytical studies for determining quality criteria. The guidance itself recommends using orthogonal techniques and not relying on one analytical method when analyzing biosimilars, notes Shakespeare.

“For example, consider analyzing the charge profile of a sample using ion exchange on a UHPLC as well as by using cIEF on a capillary electrophoresis instrument. Both techniques will show the difference in charge profiles between the innovator and biosimilar, but when a manufacturer is gathering evidence of the biosimilarity of their molecule compared to the innovator, there is strength in using multiple techniques that incorporate different instruments,” Shakespeare says.

Meanwhile, BioAgilytix, a bioanalytical lab service provider, encourages biosimilar developers to engage with their chemistry, manufacturing, and controls (CMC) team because it is important to discuss the analytical characterization of both the biosimilar and innovator biologic and note any observed differences. The company has pointed out that differences observed in an anti-drug antibody (ADA) assay, for example, “may reflect previously undiscovered analytical differences between the biosimilar and the originator” (2). It is advisable that a discussion be held with regulatory authorities if there are unexplained differences between the biosimilar and the innovator biologic, and that these discussions occur prior to moving on to the next stage of biosimilar drug development.

BioAgilytix supports a single assay approach, such as an ADA assay or antigenic assay, to support biosimilar development. When developing an ADA assay, the company advises building it in a systematic, stepwise manner. This will support the build-up of knowledge needed to instill confidence in the assay’s ability to detect antibodies against the biosimilar and the innovator biologic. To that end, the focus should be on reducing the risk for potential variability and developing a meaningful interpretation of data derived from the assay (2).

Draft Guidance for Industry: Development of Therapeutic Protein Biosimilars: Comparative Analytical Assessment and Other Quality-Related Considerations

 (CDER, CBER, May 2019).
2. T. Lester, “Questions on Immunogenicity Testing for Biosimilars Answered: Part 1,” 


Vol. 33, No. 7
July 2020
Pages: 32–33, 47

When referring to this article, please cite it as F. Mirasol, “Analytical Assays Determine Biosimilar Product Quality,” 

Articles

Features

The use of analytical assays is crucial for determining that biosimilar critical quality attributes remain on point.

Using Analytical Assays to Ensure Biosimilar Quality

Gene therapy is arguably the fastest developing therapeutic modality in biopharma. Following hot on the heels of classical biologics such as monoclonal antibodies (mAbs), an increasing number of gene therapies are entering clinical trials and receiving marketing approval. The development of gene therapies has taken longer than mAbs because of various reasons, not least of which were early setbacks and the need for more sophisticated delivery systems. While biologics such as mAbs can be administered directly to patients for therapeutic action, gene therapies tend to require delivery systems to safely transport the corrective nucleic acids to the required site of action. These delivery systems are most commonly modified viral vectors. The inclusion of vectors in the final drug product can introduce greater complexity to the processes required to manufacture gene therapies compared with those typically used for classical biologics like mAbs.

The biomanufacture of both mAbs and gene therapies requires the use of cellular expression systems, which produce the protein of interest for the final drug product. For mAbs, these systems can originate from various species, such as Chinese hamster ovary (CHO) or Escherichia coli (E. coli) cells. For gene therapies, it is much more common to use human cells, such as human embryonic kidney 293 (HEK-293), HeLa, or A549 lung epithelial carcinoma (A549) cells. To derive the final drug product, the product from the cells must be purified, using a series of steps where impurities are detected and monitored until their concentrations are low enough in the final drug product to meet regulatory requirements. An important class of impurities are host cell proteins (HCPs) (i.e., proteins from the cells used to express the protein of interest). HCPs need to be carefully controlled throughout the biomanufacturing process as a critical quality attribute (CQA). This is because HCPs can significantly affect the quality, efficacy, and safety of the gene therapy product, for example, by inducing or enhancing immunogenicity. Apart from process-related impurities like HCPs, product-related impurities like that derived from viral vectors also need to be carefully controlled.

Historically, the preferred method for analyzing HCPs and other impurities has been ligand-binding assays (LBAs), due to their speed, sensitivity, and ease of use. However, despite LBAs-namely enzyme-linked immunosorbent assays (ELISAs)-being the main regulatory compliant method in current use, there has been a steady increase in industry regulators requesting that submissions include data detailing HCP quantification obtained with methods orthogonal to ELISA. The approach that is rapidly emerging as the method of choice for many biopharmaceutical companies is liquid chromatography (LC) coupled with mass spectrometry (MS). In response to this growing need, Alphalyse and SCIEX have developed new LC-MS methods for HCP characterization of gene therapies and other new complex biologics (1, 2). These improved methods and workflows address key shortcomings posed by ELISAs in the analysis of HCPs for gene therapies. There are several main issues that are repeatedly encountered by industry players with the application of ELISAs for HCP analysis during the development and manufacture of gene therapies.

The common issue that frequently arises is with the specificity of the commercially available ELISAs. During the cell-based manufacture of various types of gene therapies, there exist numerous sources of potential residual contamination. Mammalian expression systems commonly used to generate gene therapy products, such as those involving HEK-293 cells, introduce a multitude of impurities that are secreted along with the product of interest into the cell culture medium. These impurities can include nucleic acids (i.e., DNA, RNA) and lipids, along with HCPs and other cellular material. Moreover, the media can also include other proteins that have been added to improve and expand its function, such as, for example, bovine serum albumin (BSA)-a common nutrient supplement for cultured cells-and benzonase-a nuclease enzyme that digests and removes unwanted DNA fragments. The biomanufacture of viral vectors as part of the final drug product is another source of impurity. Empty and partially filled viral capsids are product-related impurities that need to be removed from the final drug product because they can have an impact on the safety, efficacy, and immunogenicity of the therapy (3). Furthermore, the product of interest is often engineered to be human. The bioprocess of gene therapies can thus include impurities originating from multiple, diverse species, such as human from the HEK-293 cells, bovine from the BSA, viral particles from the vector system, and bacterial debris from the benzonase produced by E. coli. These contaminants from so many different species can be very difficult to detect using a single ELISA, particularly commercially available ELISAs.

Beyond the heterogeneity, the sheer number of different proteins and other impurities poses a challenge. There are potentially thousands of protein species that would need detection. This tends to be especially true for the production of gene therapies that involve mammalian expression systems and more complex biomanufacturing steps, such as the incorporation of a viral vector, than a standard biologic. Moreover, the immunogenicity of some of the proteins, particularly human ones, could be very low, meaning that it is difficult to raise detection antibodies that provide good coverage of the spectrum of protein impurities present in a gene therapy product. Even with polyclonal antibodies, it would require the use of multiple, different types of LBA to detect anywhere near the thousands of different HCPs that may be present. A single LBA is simply insufficiently specific to detect so many proteins of different types from numerous diverse species. Another issue is that of detection sensitivity. The concentrations of the various impurities can also vary greatly, with some being present in high abundance while others are present in trace amounts. These trace impurities can be as potently important for the quality, efficacy, safety, and immunogenicity of the final product as the contaminants present in more abundant amounts. Moreover, against the background of a high-volume biological sample containing high levels of the product of interest, the detection system would need to be very sensitive and have a wide dynamic range. Again, any single commercially available ELISA would not provide the sensitivity and dynamic range needed for HCP profiling of gene therapy products.

To address these issues with commercially available ELISAs, process-specific ELISAs can be developed. However, the development of these bespoke LBAs typically takes one-and-a-half to two years. This can be too long for the development of novel gene therapies because the development of this modality tends to be fast-tracked by regulatory authorities to treat serious, life-threatening conditions. Gene therapies also tend to be one-shot-and-done curative options, the first therapy to successfully reach the market tends to be the only treatment option patients need. As such, time is of the essence when it comes to the expensive gamble of developing gene therapies.

LC–MS/MS is becoming the method of choice orthogonal to ELISAs for HCP analysis because it not only offers the ability to measure thousands of proteins practically simultaneously, it can also quantify and potentially identify these proteins. It is also a generic method that can be applied easily to a broad range of gene therapy products as well as to other biologics and new modalities. These analytical advantages make LC-MS/MS assays a beneficial choice for HCP analysis. As the need for HCP analysis using orthogonal techniques grew, Alphalyse began refining a similar assay initially developed and implemented for the analysis of vaccine products (4). This developed into what is now a detailed HCP detection analysis system. The assay involves LC to separate out the analytes in the sample before quantification using tandem mass spectrometry (MS/MS). The high-performance LC is run at a microflow gradient, which eliminates any carry over analytes as well as leverages the sensitivity phenomenon unique to low-flow gradients (2). The LC-MS/MS analytical information is then comprehensively collected using data-independent acquisition (DIA) with SWATH (7,8).

MS with SWATH acquisition occurs where a TripleTOF 6600 mass spectrometer divides the mass range into small mass windows and performs MS/MS analysis on all peptides in each window (see Figures 1 and 2) (2). As the size of the mass windows are dynamic, small windows can be used to scan areas with many peptides, and larger windows can be used to more quickly analyze areas with few peptides. Using SWATH acquisition results in low interference from the high amount of the drug substance protein because the data on the low abundance HCP peptides are acquired in data-independent mode. The proteins and peptides are identified through comparison with data stored in reference databases and ion libraries. As the SWATH acquisition records multi-dimensional data with comprehensive coverage of the HCPs present, the data can be re-analyzed repeatedly in the future. The digital records thus effectively immortalize information on each protein, facilitating the continuous improvement of the annotation of the HCP profiles alongside the updating of reference databases and ion libraries with more known compounds and their fragmentation patterns (2, 5, 6).

HCP profiling by LC-MS/MS can accelerate gene therapy development

LC–MS/MS with SWATH acquisition can help bring gene therapies to market faster, safer, and cheaper. In the development of the assay, we tested and assessed its capability to detect, quantify, and identify HCPs for the monitoring and control of purity, batch-consistency, and drug release testing. The assay is sensitive enough to detect proteins present at concentrations as low as 10 parts per million (ppm), with label-free absolute quantification (2). The assay is also able to measure proteins ranging in abundance from 10 ppm to 100,000 ppm, demonstrating a wide and linear dynamic range (2). The assay can also be used to successfully monitor changes in the quantity of individual HCPs through consecutive bioprocessing steps (1). Qualitative data on the physicochemical properties of each unique protein is also acquired, including the protein molecular weight (MW), hydrophobicity, and isoelectric point (pI) (1,2). This more detailed and holistic information is particularly useful for the development and optimization of biomanufacturing processes to ensure the clearance of specific HCPs. In terms of specificity and coverage, the assay demonstrated that it was to cover the full MW and pI range of the proteome of E. coli, a common gene therapy expression system and standard reference species (2). The assay based on microflow LC–MS/MS with SWATH acquisition method is highly precise (i.e., reproducible) and robust for HCP identification and quantification (2).

The additional information provided by LC–MS/MS over ELISAs on the absolute quantification and identification of the HCPs is particularly useful to facilitate the optimization of biomanufacturing processes. The comprehensive data on the HCPs enables the straightforward modification of existing processes to optimize the clearance of the HCPs and other impurities. Another popular application of the HCP detection analysis using this assay is for the evaluation of ELISAs. As ELISAs remain the main regulatory compliant method specified by regulatory authorities, the choice of which commercially available ELISAs to use for HCP detection can be supported by LC–MS/MS data. This can confirm that the specificity, sensitivity, and coverage of the ELISA is sufficient to meet regulatory standards. To further inform the selection of the ELISAs, LC–MS/MS analysis can and is used to characterize specific HCPs of concern. The new assay can be used for comparing biosimilar drug products with their originators, as well as for analyzing preclinical and clinical batches of gene therapy product (2).

Compared with the one-and-a-half to two years it usually takes to set up a new process-specific ELISA, this new LC–MS/MS assay is quick and simple to apply for each new biologic product. It typically takes six to eight weeks to optimize and establish the assay, from the start of the project to the receipt of a regulatory notice of verification. The samples can then be analyzed on an ongoing basis. The substantially shorter timeframe and improved utility of the data acquired indicates that the adoption of this LC–MS/MS assay with SWATH acquisition could accelerate the development, marketing approval, and commercial biomanufacturing of novel gene therapies. This acceleration will likely be further enhanced as advances are continually made to the assay.

Anticipating future regulatory requirements

The biopharma industry is shifting towards including HCP data from orthogonal methods to LBAs, primarily from MS assays. This trend is observed in the industry as companies increasingly request detailed HCP detection analysis using this new assay based on microflow LC–MS/MS with SWATH acquisition. It is expected that by preparing such detailed HCP data, which can continue to be better annotated over time, the chances of obtaining regulatory approval for clinical trials and marketing are improved. Moreover, it is an important component in the risk management strategies for many large companies investing in the costly and competitive development of new gene therapies. Implementing measures to minimize the need to re-run analyses later in the drug development cycle not only saves time, but could also prove more cost efficient.

“Identification and Absolute Quantification of Individual Host Cell Proteins by SWATH LC–MS,”

 presentation at the Well Characterized Biologics Conference (Rockville, MD, Oct. 24–26, 2018).
3. T. Li, Gao, et al., 

“Determination of Full, Partial, and Empty Capsid Ratios for Adeno-Associated Virus (AAV) Analysis,”

 SCIEX Technical Note, sciex.com, accessed April 2020.
4. S. Heissel, et al., 

“Host Cell Protein Analysis by Microflow-LC High Resolution SWATH-MS of Vaccine Samples Under Development,”

 poster presented at The American Society for Mass Spectrometry (ASMS) Annual Conference (San Antonio, TX, June 5–9, 2016).
5. SCIEX, “

SWATH Acquisition, Ensuring Nothing is Missed,”

 sciex.com, accessed April 2020.
6. C. Carapito, 

“Dual Data-Independent Acquisition Method using SWATH Acquisition–MS for Host Cell Proteins (HCP) Profiling and Absolute Quantification of Key Impurities during Bioprocess Development,”

 SCIEX Webinar, sciex.com, accessed April 2020.

Ejvind Mørtz, PhD, is co-founder and COO of Alphalyse. He has more than 20 years of experience from several biotech companies, including the development of protein analysis methods in research and chemistry, manufacturing, and control development of protein biologics, and business collaborations with contract manufacturing organizations/contract researche organization.

This method is expected to help bring gene therapies to market faster, safer, and cheaper.

A Novel Method for Host Cell Protein Analysis

MilliporeSigma plans to build a new, $20-million laboratory facility in Buchs, Switzerland, to support its rapidly growing reference materials business, the company announced in a July 15, 2020 press release. The company anticipates that about two dozen jobs will be created with this expansion.

 Buchs building is scheduled to be done by December 2021, and the move to the facility is planned for early 2022. The facility will include laboratory and office space aimed at fostering a collaborative environment. The new facility will offer a more efficient working environment for current and future employees in R&D, analytical production, and quality control. The laboratory area can be adapted to new requirements in the future, including increased site capacity.

"Researchers in testing labs around the world use MilliporeSigma’s reference materials every day to help generate accurate, reliable analytical results in pharmaceutical, environmental, and food and beverage analysis,” said Jean-Charles Wirth, head of Applied Solutions at MilliporeSigma, in the company press release. “This new laboratory will allow us to continue to drive innovation in diagnostics and testing and expand our research and development of analytical standards.”

News

The investment will allow for the expansion of analytical R&D efforts and will support the company’s reference materials business.

MilliporeSigma Invests $20 Million in New Switzerland Laboratory

MOBILion Systems, a United States-based instrumentation company specializing in biomarker discovery, diagnostics, and therapeutic development, now offers its new ion mobility separations technology, structures for lossless ion manipulation (SLIM) for commercial use. SLIMS technology offers efficient separation, identification, and analysis of the most challenging, clinically significant molecules that other instruments fail to detect, the company announced in a July 22, 2020 press release highlighting its first-half 2020 progress.

“In the first half of 2020, our teams continue to implement [this technology] successfully, and the first-use cases of the SLIM technology are proving to be disruptive,” said Melissa Sherman, CEO of MOBILion Systems, in the company press release. “Despite COVID-19 we have hit all milestones and are being tapped by our beta users who want to use SLIM for COVID research. SLIM’s high resolution unlocks small structural differences that impact the research of the virus, the disease and its potential therapeutics.”

The SLIM technology can be integrated with liquid chromatography–mass spectrometry (LC–MS) workflows to provide more robust analytical information. For some applications, it can replace LC to provide higher resolution, speed, ease of use, and greater instrument uptime.

New Technology

The company’s SLIM technology has been a part of the company’s accelerated growth by offering enhanced R&D analytics.

MOBILIon’s SLIM Technology Enhances R&D Analytics

Polyplus-transfection, a France-based biotechnology company focused on gene and cell therapies, has launched a current good manufacturing practices (cGMP)-compliant residual test for its PEIpro product portfolio, transfection reagents designed for process development, pre-clinical, clinical, and commercial lentivirus and adeno-associated virus (AAV) production for cell and gene therapies. The company is also launching dual sourcing for its PEIpro-GMP product for late-stage and commercialization of cell and gene therapies, Polyplus announced in a July 20, 2020 press release.

The cGMP-compliant test for its PEIpro product range is designed to detect residual transfection reagents during the production of advanced therapeutic medicinal products (ATMPs). cGMP guidelines specify that manufacturers should be aware of both the residual levels of raw materials used in drug products and the significance of their presence. These regulations ensure manufacturers reliably determine residual raw material levels and maintain reproducible safety of ATMPs for patient administration. The Polyplus test is able to detect and quantify residual PEIpro reagent throughout the ATMP manufacturing process. It can thus be used as a quality control measure for product release. The test can be adapted for each ATMP to ensure the lowest limits of detection.

In parallel, Polyplus has introduced dual sourcing for its reagent, which will mitigate the risks to the ATMP industry. The dual sourcing approach enables Polyplus to source its PEIpro-cGMP product from two distinct subcontracting manufacturing plants and enables the company to ensure sole responsibility and control of the process. ATMP manufacturers are then able to have Polyplus as a single point of contact while benefiting from increased production capacity and shortened lead times.

“As increasing numbers of ATMPs and gene therapies move through late-stage trials and commercialization, demand on cGMP reagent supply continues to increase. The complexities of ATMP therapies mean that Polyplus has to increase its services to the sector alongside increasing its product portfolio,” said Claire Wartel, PhD, director of quality and compliance, Polyplus-transfection, in the company press release. “By offering PEIpro residual testing and dual sourcing of PEIpro-cGMP to the ATMP market, this enables sector maintenance of regulatory compliance in addition to the lowering of manufacturing and supply risks. Polyplus will continue to work with the ATMP sector to develop further tests and services for therapies for patients.”

The company has launched a residual GMP-compliant test and dual sourcing opportunity designed to increase compliance and reduce risks for ATMP manufacture.

Polyplus-transfection Launches Residual Test for Transfection Reagents 

Primera Technology introduced its Signature Slide and Cassette Printers with integrated PTLab software for increasing lab efficiency and reducing the risk of specimen misidentification.

According to a July 13, 2020 press release, the printers can print text, graphics, logos, and 2D bar codes directly onto slides and cassette. The devices also include:

Reduced inventories and purchasing activity

Thermal-transfer printing at high-resolution, 300 dpi

The printers can print text, graphics, logos, and 2D bar codes directly onto slides and cassette.

Primera Technology Introduces Signature Slide and Cassette Printers

Rigenerand, a Modena, Italy-based biotech company specializing in medicinal products for cell therapy applications, has begun development of a novel 3D bioreactor and pipeline specifically designed for the incremental production of extracellular vesicles exosomes (EVs) in a current good manufacturing practices (cGMP) environment. The development will be supported with part of a EUR 4.4-million (US$5.1-million) grant from Horizon 2020 for the H2020-EU.1.2.2.-FET Proactive program, awarded to the Biogenic Organotropic Wetsuits project. This project comprises a consortium of companies, including Rigenerand.

The Biogenic Organotropic Wetsuits project involves 11 organizations across seven countries in Europe. Rigenerand will contribute its combination of expertise, technology, and innovation focused on biomaterials, prototypes production, and industrialization of cell-culture devices and 3D cell cultures to the consortium, the company said in a July 8, 2020 press release. Rigenerand will start developing novel ways of improving current EV cGMP production technologies, which will ultimately aim to produce EV clinical batches to be used as drug substances for cell-free therapy-based medicinal products.

The project is due to start in the fourth quarter of 2020. By the second quarter of 2021, Rigenerand plans to have developed the isolation and expansion of a human cellular source (EDT-MSC) suitable for the production of immune-ghost EVs as well as the separation and production of suitable amounts of EV for characterization and pre-clinical application.

“This project will contribute to the urgent medical need of increasing EV production. It will enable Rigenerand to develop pre-conditioning approaches that are critical to providing solutions to these challenges,” said Massimo Dominici, scientific founder, Rigenerand, in the company press release. “The activity for this consortium will also enable Rigenerand to develop platforms for new therapeutic solutions, covering a range of clinical applications, including oncology and regenerative medicine to medical devices.”

The Rigenerand 3D bioreactor for EV production will be based on its proprietary VITVO technology. VITVO is a handheld bioreactor, with an integrating scaffold for the establishment of a 3D cell culture model. It is already used for highly predictive pre-clinical testing and has been specifically designed to offer an increase to the market in usability, sizing, closed system performance, and flexibility for 3D cell culture technologies.

The company has begun development of a 3D bioreactor for the cGMP production of extracellular vesicle exosomes.

Rigenerand Develops 3D Bioreactor for Extracellular Vesicle Exosomes Production

Intravacc, a Netherlands-based vaccine research and development company, announced the publication of a study in 

 about the alternatives to the use of laboratory animals in vaccine quality control. The study was led by the Leiden Academic Center for Drug Research (LACDR) at Leiden University.

According to a July 23, 2020 company press release, Intravacc and LACDR reproduced vaccines in test tubes and treated them with cathepsin S (CS), an enzyme that breaks down vaccines and pathogens into fragments that are recognized by immune cells. The study found that, instead of CS slowing down the breakdown of the vaccine as the researchers expected, the opposite occurred, and the researchers were able measure vaccine inactivation by quantifying the formation of vaccine fragments during the breakdown by CS.

The team concluded that it may be possible to replace animal testing in the future for vaccines using this inactivation, the press release said.

“As a result of the Covid-19 pandemic, some 400 therapies and vaccines against this virus are currently being developed worldwide,” said Dr. Jan Groen, CEO of Intravacc, in the press release. “This affects the number of laboratory animals used in studies. Intravacc considers limiting the need for animal testing to be particularly important and plays an important role in the development of alternatives for this vaccine-related research. This study shows the progress we are making on this front.”

A study in published in Scientific Reports showed that researchers were able measure vaccine inactivation by quantifying the formation of vaccine fragments during the breakdown by CS.

Intravacc and Leiden University Develop Possible Replacement for Animal Testing in Vaccines

LabVantage Solutions, a laboratory informatics solutions and services provider, announced that its newly released 8.5 edition of its laboratory information management system (LIMS) platform now offers a fully integrated scientific data management system (SDMS).

Along with guaranteeing security and control against data loss or modification, the SDMS system works to automatically collect all relevant lab-generated data for use by laboratory staff, a July 8, 2020 company press release said. The additional functions of the LabVantage SDMS include:

Capturing externally generated data from instruments, data sources, incoming emails, and other file sources

Reducing bottlenecks by deploying distributed file collectors

Using a centralized LIMS-controlled file system, database, or Amazon web service (AWS) cloud for storage

Finding and linking back to files for rapid access and restoration for further use

Creating graphical Talend-based parsers to extract and process data.

“During these challenging times, we are especially proud to release LabVantage 8.5, which represents a major advance reflecting our commitment to making the advantages of digital transformation fully accessible to our customers,” said John Heiser, CEO of LabVantage, in the press release. “LabVantage 8.5’s fully integrated SDMS is specifically designed for the LabVantage platform and provides major benefits to laboratory managers, with the aim of making their operations more efficient, productive, and secure. Our user-friendly, all-in-one SDMS offers seamless, continuous data integrity. Importantly, it allows managers to secure all of their data, not just the final results.”

Along with guaranteeing security and control against data loss or modification, the SDMS system works to automatically collect all relevant lab-generated data for use by laboratory staff.

LabVantage Adds an SDMS to the 8.5 Edition of its LIMS Platform

Pharmaceutical Technology In the August 2020

Pharmaceutical Technology's In the Lab eNewsletter 08-05-2020