Extractable and leachable (E&L) studies have become an integral part of any drug development program and serve to ensure compatibility between the product and its packaging. While such studies are now generally well defined in terms of experimental design and data evaluation, these have had to adapt to the ever-increasing diversity of therapeutic products. There are, for example, some unique challenges in the analysis of biopharmaceutical products as a result of certain formulation components, as well as the complexity of the therapeutic itself. This article describes a number of challenges and solutions in the E&L evaluation of biopharmaceuticals and highlights a selection of those encountered with emerging cell and gene therapy (CGT) products.

The risks posed by leachables on product quality and patient safety are an ongoing challenge for the pharmaceutical and biopharmaceutical industries. Continued investigational development in this area is primarily driven by a number of factors, including: the increased adoption of single-use technologies; progress in the medical device field, particularly combinational devices; expanded directives from industry regulators; and advances in analytical technologies.

Extractables (1) are defined as compounds that migrate from any product-contact material (including elastomeric, plastic, glass, stainless steel, or coating components) when exposed to an appropriate solvent, under exaggerated conditions of time and temperature. To study materials at risk of potential leachable components, extractions of the contact material are performed, generally using aggressive conditions including acidic, basic, organic, and aqueous solvents. These conditions are not intended to distort the material under investigation but do attempt to mimic the drug formulation. The resulting extracts are usually analyzed using a combination of analytical techniques such as liquid chromatography–mass spectrometry (LC–MS), gas chromatography–mass spectrometry (GC–MS), and inductively coupled plasma–mass spectrometry (ICP–MS) that can qualitatively and quantitatively evaluate both the organic and inorganic components, or “extractables”. Such studies are typically performed to represent a worst-case scenario and may provide an indication of possible leachable components as well as facilitate material selection during early risk assessment.

Leachables (1) are compounds, often identified during extraction studies, that migrate into a drug formulation from any product-contact material (elastomeric, plastic, glass, stainless steel, or coating components). This leaching happens as a result of direct contact under normal process or accelerated storage conditions, such as elevated temperature. However, if the leachable interacts with the drug product (as may be the case particularly with biotherapeutics) or packaging materials, new components can be generated that are known as “secondary leachables”. The complexity of many formulations, often containing a variety of buffers, surfactants, fillers, and other excipients, can lead to the formation of secondary leachables only identified after long-term stability studies.

E&L evaluation of traditional pharmaceutical medicines is now well established and may be undertaken with a certain predictability in experimental design and even outcome. In contrast, however, certain excipients used in the formulation of some biotherapeutics present unique analytical challenges for leachable determination. Among these excipients are the commonly used nonionic surfactants, such as polysorbate 80 (Tween 80), which constitute a mixture of fatty acid esters of polyoxyethylene sorbitan. These have an extremely low ultraviolet (UV) response but are highly responsive to ionization methods commonly used in LC–MS (2). This often leads to significant interference, either by co-elution and/or suppression during analysis, raising the real possibility of contaminants remaining undetected. Extraction of these surfactants prior to analysis is a possible means of mitigation but is often not highly effective due to the surfactant’s inherent heterogeneity and limited extraction efficiency. Care must also be taken in the application of such an approach since the leachable may be co-extracted with the surfactant.

In the growing area of CGT, there are currently no specific guidelines for E&L studies. However, such products have some unique challenges, such as the use of “ancillary materials”. These materials are used during manufacturing but are not intended to be present in the final product. They are often themselves complex biological materials, such as recombinant insulin and human serum albumin, used as media supplements, and collagen, used as a scaffold in implantable devices (3). Since these materials can potentially interact with and be affected by leachables, in the same manner as the biotherapeutic itself, the choice of materials and controls must be carefully considered to avoid adverse consequences for the manufacturing process.

The evaluation of biotherapeutics for leachables presents many challenges, some of which are somewhat unique to this type of product. For example, the biopharmaceutical itself may interfere in the testing regimen due to its high concentration, heterogeneity, and/or high detector response. A potential solution is to remove the biotherapeutic prior to analysis, but the procedure must be very carefully considered because associated leachables may be unknowingly removed and/or additional extraneous components may be introduced.

The major concern of leachables in pharmaceutical products is their specific toxicity and/or genotoxicity. The additional issue with biotherapeutics is the potential for interaction between the structurally complex biopharmaceutical itself and any leachable components. Compared to traditional pharmaceuticals, proteins and other biotherapeutics are often significantly less stable, due to their strong dependence on physico-chemical and conformational structural properties. These larger molecules often have both hydrophilic and hydrophobic regions, with many reactive sites for possible leachable binding, potentially resulting in the loss of activity via mechanisms such as unfolding, truncation, aggregation, and precipitation. Consequently, the adverse effects of extractables and leachables can be more significant and more difficult to assess on biopharmaceutical products than small molecules. Acrylic acid, for example, a leachable from syringes, has been shown to react at three different sites within a protein structure, resulting in potential changes in charge and hydrophobicity with consequential effects on the therapeutic properties of the protein-based drug (4).

As the manufacture of many biotherapeutics involves cellular-based processes, the presence of certain leachable components can have unfavorable consequences not only on the final drug product but also on the production process itself. Productivity can certainly be adversely affected by leachables that result in cellular toxicity, leading to significant loss in manufacturing yield.

During the past decade, many biotherapeutic manufacturers have migrated to the use of disposable, single-use systems (SUS) for production, and this has now become commonplace in the emerging CGT field. Every operational phase of a biomanufacturing process that uses SUS technology is at risk of leachable contamination. This contamination can be introduced as a result of contact and interaction with different materials, such as polymers, metals, and additives used to construct holding tanks, films, bags, tubing, and filters. This concern extends to containers for buffers, media, water for injection, and other liquids required for manufacturing. The possibility of leachable accumulation may also arise since concentration stages are often used in the manufacturing process.

The majority of biopharmaceuticals are injectable products and, therefore, contaminants enter the bloodstream directly, without the potential for depletion in the gut as with oral medications. In addition, many biotherapeutics, particularly monoclonal antibodies, are dosed in high concentrations (multiple mg/mL) and are produced in liquid, with complex formulations. These drugs can be stored for long periods of time, providing higher risk of leachable adulteration of the product than the solid dose forms common with traditional pharmaceuticals.

Certain products developed in the CGT area can present further challenges for E&L evaluation. Since the quantities of product can be relatively low, the ratio of contact material surface to the drug volume is very high, increasing the potential for leachable contamination. This may be further exacerbated in those treatments that are essentially customized, with each patient receiving the entire product dose.

Risk assessment in terms of E&L is now an integral part of any product development strategy. This is of particular importance for CGTs, which should not only evaluate the likely compatibility of contact materials used in manufacturing, but also include an experimental study designed to provide an accurate assessment of the extractable and leachable profiles.

While many biotherapeutics are produced as liquid formulations, lyophilized forms of drug product are also commonplace, as part of efforts to improve stability during storage. Interestingly, the FDA Center for Drug Evaluation & Research, the FDA Center for Biologics Evaluation & Research, and the United States Pharmacopeial Convention (in 

 <1664> [5]), describe sterile powders for injection as being “low risk” for leaching, despite some evidence to the contrary. It has been demonstrated, for example, that several semi-volatile organic compounds originating from butyl rubber stoppers—part of the primary packaging system—may leach into a lyophilized product but not a liquid formulation stored in the same packaging (6). The explanation for this finding lies in the hydrophobicity of the leachables, rendering them very poorly soluble in aqueous media but able to diffuse from the stopper. This is perhaps facilitated by their volatility and the nature of the drug product.

There are multiple types of material used in the manufacture and storage of biopharmaceuticals, and most may be classified as either glass, metal, or synthetic polymer/elastomer. The latter are recognized as the most significant sources of leachables, often resulting from monomers/oligomers of the polymer itself, as well as catalysts, initiators, additives (e.g., lubricants, antioxidants, antitack, and antistatic agents), adhesives, anchoring agents, adhesive resins, colors, fillers, and degradation products of almost all these components. The diversity and nature of these potential contaminants and their often-unpredictable interactions with formulated biopharmaceuticals pose significant challenges to analytical assessment of the drug products and process materials. There are now significant guidance documents available from regulatory authorities, industry organizations, and researchers providing at least some support in the design and rationale for suitable experimental approaches.

For biopharmaceuticals, inorganic leachables (generally associated with glass and metal) are of particular concern because of their potential impact on the stability and efficacy of the drug itself. A metal commonly used is stainless steel, which has found applications in manufacturing, shipping, and storage, and is known to leach iron, chromium, and nickel, particularly if the steel is unpassivated (7). Trace levels of metal ions can potentially cause protein degradation by mechanisms such as protein oxidation and fragmentation. The latter occurs because of the metal-binding propensity of aromatic amino acids such as tryptophan, tyrosine, and histidine. Certain metal–protein binding is also known to induce secondary and tertiary structure changes resulting in the formation of insoluble protein aggregates (8).

Glass has also been demonstrated to be a source of certain metal leachables. For example, manganese and iron oxide—used as coloring agents—are present in Type I amber vials as are zinc and

barium oxides. Higher levels of manganese and iron have been shown to leach from amber glass compared to clear glass. Meanwhile, sterilization techniques, such as steam autoclaving and gamma irradiation, may also lead to the presence of metal ion contamination (9).

Some biotherapeutic excipients, and the drug itself, may also play a significant role in facilitating the extraction of metals from contact surfaces. Certain proteins and chelating agents, such as ethylenediaminetetraacetic acid, have been shown to increase the quantity of metals such as aluminum, calcium, and iron from plastic container systems. Similarly, buffers such as phosphate and lactate can also increase the metal leaching from both metal and glass surfaces.

The area of E&L investigation has developed rapidly over the past decade, resulting in technological and regulatory advancement. Ultimately, this has led to safer medications. However, developments in biopharmaceuticals and specifically in CGT are likely to present further demands on expertise in areas of manufacturing and analytics.

It is clear that a single, common strategy, for monitoring and/or eliminating leachable contaminants will never be sufficient to address all situations. Instead, engineers and investigators must be always prepared for the unexpected.

1. D.B. Lewis, “Current FDA Perspective on Leachable Impurities in Parenteral and Ophthalmic Drug Products,” Presentation at AAPS Workshop on Pharmaceutical Stability—Scientific and Regulatory Considerations for Global Drug Development and Commercialization (Washington, DC, Oct. 22–23, 2011).
2. J. Fukuda, et al., 

 B 953–954, pp. 68–72 (2014).
3. J. Solomon, et al., 

 18 (1) 1–12 (2016).
4. D. Liu, et al., 

 <1644>, “Assessment of Drug Product Leachables Associated with Pharmaceutical Packaging/Delivery Systems,” 

, first supplement (Rockville, MD, Aug. 1, 2015).
6. S.A. Zdravkovic, 

 107 (11) 2837–2846 (2018).
7. L. Allain and Q. Wang, 

 10 (3) (2007).
8. J.L. Cleland, M.F. Powell, and S.J. Shire, 

 10 (4) 307–77 (1993).
9. D. Bohrer, et al., 

Mark Rogers, PhD, mark.rogers@sgs.com, is the global scientific director for the SGS Health Science Drug Development group.

Articles

Features

Establishing extractable and leachable studies for complex biotherapeutic molecules poses new challenges for analytical assessment.

Preparing for Unexpected Issues in E&L Studies for Biopharma

Over the past two decades, the biopharmaceutical industry, along with the associated regulatory authorities and analytical instrument suppliers, have progressed toward clearer identification of the attributes that are critical for ensuring the quality of biopharmaceuticals and how to measure them efficiently. 

The need to understand and predict these molecules’ behavior drives a requirement for new informational insights. This article puts the spotlight on infrared spectroscopy and recent innovations, including the introduction of Microfluidic Modulation Spectroscopy.

Holly Lombardo*, HLombardo@redshiftbio.com, is product manager, and Matthew McGann is marketing manager; both are at RedShiftBio.

*To whom all correspondence should be addressed.

A recent innovation in infrared spectroscopy, Microfluidic Modulation Spectroscopy has been shown to improve characterization of secondary protein structure.

Improving IR Spectroscopy as a Tool for Biopharmaceutical Analysis

Thermo Fisher Scientific has introduced Ion Torrent Oncomine immune repertoire assays, a new suite of next-generation sequencing (NGS) assays that are designed to detect potentially malignant clones of T cells and B cells, key players in immune response. The pan-clonality assays use proprietary Ion AmpliSeq technology to target multiple parts of B- and T-cell immune receptors. This is done using a single reaction with ultra-high sensitivity, which increases the probability of detecting malignant clones and decreasing the time to results, Thermo Fisher announced in a July 19, 2021 press release.

Detection of distinct receptor DNA sequences can inform diagnostic, prognostic, and therapy development studies, which can illuminate the progression of lymphoid cancers, including leukemias, lymphomas, and diseases such as multiple myeloma. Using NGS technology, detection sensitivity can be increased—compared to traditional testing methods, such as flow cytometry or quantitative polymerase chain reaction.

One of the challenges of current NGS-based immune repertoire analysis has been the need to run multiple secondary tests because of the high test failure rates of a typical single NGS assay, the company explained in the press release. The new Oncomine pan-clonality assays overcome this challenge by uniquely targeting multiple parts of the receptor to reduce the need for secondary tests and increase the probability of a single assay detecting a potentially malignant clone.

"Genomic testing can make a significant impact on researching and understanding these [blood] cancers—but we need to bring more testing to in-house laboratories where results can be made available quickly to healthcare professionals," said Garret Hampton, president of clinical next-generation sequencing at Thermo Fisher Scientific, in the press release. "The new Oncomine immune repertoire assays unlock the power of NGS to help researchers rapidly obtain a tremendous amount of insight about a lymphoid cancer sample, including its prevalence and an understanding of how it is evolving."

"With the ability to sequence multiple targets in a single reaction, our team can now get much more information from one assay when compared to single-target NGS tests," said John DeCoteau, medical director, University of Saskatchewan, Advanced Diagnostic Research Laboratory, in the press release. "Using the Oncomine BCR Pan-Clonality Assay, we were able to achieve a positive clonality detection rate of 93 percent in a cohort of multiple myeloma samples, without the need for secondary testing. That provides a remarkable improvement for laboratory efficiency and allows us to get results faster."

New Technology

Thermo Fisher Scientific has added a new suite of immune repertoire assays that offers high detection rates of malignant clones, allowing researchers to better assess blood cancers.

Thermo Fisher Expands Hematology Oncology NGS Portfolio

On July 27, 2021, Byondis, a Netherlands-based clinical-stage biopharmaceutical company, and Glycotope, a Germany-based biotechnology company specializing in antibodies that target tumor-specific-carrying carbohydrate structures, have entered into a platform access agreement to discover and develop antibodies that target specific tumor-associated protein/carbohydrate combined glyco-epitopes (GlycoTargets).

Under the agreement, Byondis has gained exclusive rights to evaluate and develop antibodies against selected novel GlycoTargets and has the option to in-license these antibodies for development as antibody-drug conjugates (ADCs). Financial terms were not disclosed.

Glycosylation is strongly altered in cancer cells, which reflects the drastic changes in tumor metabolism. This alteration, in turn, results in new, highly tumor-specific GlycoTargets. Specific antibodies against these GlycoTargets might be highly cancer specific and have the potential to target a broad range of oncology indications, the companies stated in a press release.

Formulating the antibodies into ADCs can improve their cell-killing capability. But while earlier generation ADCs improved targeting and cell-killing, they were unstable in the bloodstream, which led to premature release of the cytotoxic payload that impacted healthy tissue and narrowed the therapeutic window. Byondis’s next generation ADCs are designed to be highly stable in circulation and carry an intricate, inactivated, and potent cytotoxic drug that rapidly self-destructs if prematurely released, thus limiting damage to healthy tissue and improving the therapeutic window, according to a Byondis press release.

“Selective tumor targeting is key to maximiz[ing] the potential of our linker-drug platforms to generate effective ADCs with low systemic toxicity. The antibodies generated by Glycotope are designed to deliver very high tumor selectivity. We are excited to join forces and align our highly complementary technologies to provide effective and safe ADCs for the benefit of cancer patients,” said Byondis Chief Scientific Officer Wim Dokter in the press release.

“This agreement with Byondis highlights the potential of our GlycoTarget discovery and technology platform,” said Henner Kollenberg, managing director of Glycotope, in the press release. “GlycoTargets are a unique class of highly tumor-specific antigens that offer reduced on-target/off-tumor toxicities. We believe that combining antibodies against these GlycoTargets with Byondis’ linker-drug platform has great potential to produce novel treatments for people suffering from diseases for which no adequate medical treatment option has been found so far.”

News

Byondis and Glycotope have formed an agreement to discover and develop antibodies that target specific glyco-epitopes.

Byondis and Glycotope Partner on Discovery and Development of Glyco-Epitope-Targeting Antibodies

Summa Equity has acquired a majority stake in Axion BioSystems, a US life science tools business specializing in microelectrode arrays (MEA) and impedance technology for single-cell analysis. Axion provides an integrated technology platform for cell activity monitoring, typically used by academia or biopharma customers for non-invasive, real-time study of cells. Axion’s technology was of particular interest to Summa with regards to cell and gene therapy (CGT), which could be instrumental in curing various acquired and genetic diseases.

“Summa has been focused on solutions in the CGT market for a long period of time, due to its alignment with our strategy and its significant potential for growth,” said Tommi Unkuri, a partner at Summa Equity, in a press release from July 16, 2021. “Axion BioSystems impressed us with its … scalable technology, which gives both academics and biopharma organizations the ability to go much deeper in their understanding of cell behavior, enabling faster, more accurate and more cost-efficient drug discovery, development and quality control.”

“We have been highly impressed with Summa’s understanding of the life sciences market, and belief in the potential of our technology to accelerate innovation in many areas of pharmaceuticals and healthcare,” said Tom O’Brien, CEO and president, Axion BioSystems in the press release. “We believe that they will bring tremendous value in helping us to increase our commercial capacity, reach, and market awareness to scale up significantly and have an even greater impact in the years to come.”

Summa Equity has acquired a majority stake in science tools business Axion BioSystems with the intent of further entering the CGT market. 

Summa Equity Acquires Majority Stake in Axion BioSystems 

The innovation deal between 908 Devices and Transcenta in mid-2018 has materialized into concrete improvements in workflow efficiency. By utilizing 908 Devices’ technologies at their manufacturing facility in Hangzhou, China, Transcenta has been able to facilitate their biopharma analytical workflows across large molecule characterization and formulation process development.

908 Devices’ ZipChip enables the rapid measurement of critical quality attributes (CQAs) of intact monoclonal antibodies (mAbs). By utilizing this technology, Transcenta was able to characterize intact mAbs and determine what was causing increases in acidic and basic variants when placed under stress at high temperatures. The antibodies were similarly analyzed in their intact state under native conditions as well as their subunit and peptide levels. Ultimately, the analysis helped determine heterogeneity speciation and identification in under ten minutes, which resulted in a workflow that was 100 times faster for Transcenta.

“Technologies offered by 908 Devices enabled us to quickly obtain high-quality analytical data to help accelerate development of innovative biologics to ultimately reach patients sooner. Our recently published results demonstrate the positive impact,” said Christopher Hwang, chief technology officer of Transcenta, in a press release from July 22, 2021. “ZipChip is a capable platform for multiple CQA assays, and, for charge variant analysis, it is very straightforward to get separation and identification information with minimal method development needed for different types of proteins, especially for the analysis in native conditions. The sample preparation is simple and straightforward, and the sample requirements are at nanogram level.”

In addition to the ZipChip, Transcenta also found success with 908 Devices REBEL technology. REBEL is designed to accelerate media and process development, both of which are critical in the development of highly intensified continuous cell culture perfusion platforms. REBEL was instrumental in getting Transcenta to a point where its volumetric production rate rose to more than 6 g/L per day. More specifically, it was crucial in their amino acid analysis.

“REBEL can analyze a large number of samples in a short period of time to provide valuable data to accelerate cell culture process development,” continued Hwang. “Rather than obtaining amino acid profiles after the run is completed, REBEL can provide near real-time data for us to make process decisions while the experiments are in progress to maximize successful outcomes.”

“The pharmaceutical industry recognizes that enhanced process understanding and control lead to improved yields and predictability. Our ZipChip and REBEL products enable that understanding by making bioanalysis fast and accessible,” said Kevin J. Knopp, CEO and co-founder of 908 Devices, in the press release. “The outcomes of our collaboration with Transcenta are encouraging and serve as a beachhead for our expanding work in the Asia Pacific region. We’re proud of the successful work and discoveries developing from the use of our products in this region.”

Following a deal struck in mid-2018, 908 Devices’ ZipChip and REBEL technology have proven instrumental in facilitating Transcenta’s efficiency and volumetric production.

908 Devices and Transcenta Partnership Leads to Workflow Innovations

Ziath, a UK-based instrumentation control and information management company focused on laboratory automation, in conjunction with the University of Hertfordshire, has discovered a means of using artificial intelligence (AI) to enable discrimination between empty wells in sample tube racks from wells that may have an obscured or poorly rendered barcode.

Ziath utilizes DataMatrix barcodes to track biological and compound samples. These barcodes are laser-etched onto the underside of sample tubes, which are then stored in racks. Tube identification is then done via a barcode reader that scans the bottom of the rack and decodes it.

A problem had emerged, however, where traditional barcode readers were sometimes unable to determine the difference between an empty well and those that have hard-to-read labels. This error can happen for various reasons, such as ambient lighting, background image lasering, or simple variation in barcode lasering and material.

Alexander Beasley, PhD, from the University of Hertfordshire, used his expertise in embedded systems design to develop a scanner that circumvents these issues. By utilizing a convolutional neural network (CNN) for feature extraction of images, Beasley has been able to successfully discriminate between empty and full wells.

The new technology is expected to launch in Ziath barcoded tube scanners by the end of 2021. In the meantime, Ziath has also implemented the technology in the latest version of its DP5 control software, offering customers the opportunity to try the new technology immediately.

“The CNN I have chosen is designed to be very lightweight, allowing for quick execution,” said Beasley, in July 8, 2021 press release. “When compared to the pre-existing heuristic methods, the CNN approach was almost ten times faster to execute with virtually 100% accuracy.”

“This is just the first deliverable from our collaboration with Alexander and the University of Hertfordshire team,” said Ziath Managing Director Neil Benn in the press release. “We are expecting this project to revolutionize the way we decode DataMatrix tubes and help us produce the next generation of faster, lighter, go-anywhere tube rack readers. It’s an exciting development that will, very soon, improve sample tracking and tracing for scientists everywhere.”

Through a partnership with the University of Hertfordshire, Ziath discovers means of using AI to detect empty wells in sample tube racks.  

Artificial Intelligence Facilitates Empty Well Detection 

Avantor’s new J.T.Baker robotic tips, a series of robotic tips designed for use in robotic liquid handling and research workstations launched in July 2021, are designed for a range of applications, including genomics, cell biology, and proteomic workflows. They come in two formats: conductive and clear.

The conductive tips are designed to minimize downtime and keep operations running smoothly. Small liquid volumes can be sampled with high precision and without contamination. Additionally, the Hamilton and Tecan-type tips can be used in many robotic stations.

The clear tips are built for both sample preparation and drug discovery applications. Because these tips are transparent, users can visually recognize when the product is foaming for sampling purposes.

The J.T.Baker robotic tips are the latest in Avantor’s line of automated precision plastics, and one of the first to be announced following the company’s recent acquisition of Germany-based Ritter (and affiliates), a manufacturer of high-quality robotic and liquid handling consumables, including conductive pipette tips, in June 2021. Each batch is independently tested to make sure there are no contaminants such as DNase, RNase, amphipathic tail-anchoring peptide, and pyrogens.

“Our new, high-quality J.T.Baker robotic tips help researchers get the reliable, repeatable results required to keep critical life sciences work moving forward,” said Rohit Shroff, senior vice-president of Global Lab Products at Avantor, in a July 12, 2021 company press release. Our tips provide customers the precision and reliability they need to support their dynamic requirements.”

Avantor’s newly unveiled J.T.Baker robotic tips is the latest in their line of automated precision plastics.

J.T.Baker Robotic Tips from Avantor Offer Precision Workflow 

Generate Life Sciences announced the establishment of its good manufacturing practice (GMP) facility in La Jolla, Calif. for newborn stem cell biologics end-to-end manufacturing. This facility will add the role of a personalized therapeutics developer with in-house manufacturing capabilities to the company’s core international donor gamete, genetics service, and newborn stem cell preservation business.

With its bioreactor system, automation, and its meeting cGMP regulations, the new facility is designed to maximize bioprocessing efficiency of newborn stem cells and manufacturing at scale.

"The La Jolla GMP facility will be a crucial addition to Generate's vision for our newborn stem cell operation," said Jaime Shamonki, chief medical officer of Generate Life Sciences, in a press release. "As the largest private newborn stem cell biorepository in the world, we believe it is our duty to continually invest in new scientific applications for newborn stem cells with the goal to increase access to novel therapies for families. With this facility, we will have the in-house capability to take these stem cells through the complete process of collection, cryogenic storage, and ultimately biologics manufacturing. This will allow us to power our studies in a much more efficient and streamlined manner and help us better serve the families who have stored over 1.2 million stem cell units with us."

Mesenchymal stem cells (MSCs) can be isolated from the umbilical cord tissue and used in regenerative medicine due to their anti-inflammatory, immune-modulating, and tissue reparative properties. Researchers and scientists are targeting the use of MSCs for the treatment of conditions such as autoimmune disease, acquired neurological diseases, lung disease, and conditions related to infertility. In addition, MSCs are being used in clinical trials for cell therapy, gene therapy, tissue-engineering, and combination products.

The GMP facility is designed to maximize bioprocessing efficiency of newborn stem cells and manufacturing at scale.

Generate Life Sciences Establishes Newborn Stem Cell Manufacturing Facility

On July 7, 2021, Thermo Fisher Scientific announced its expanded gene therapy portfolio consisting of a new media panel, gene kit, and advanced purification resins to support adeno-associated viral (AAV) manufacturing. The tools aim to reduce manufacturing costs while increasing the viability of gene therapies as treatment options for patients.

The Gibco Viral Vector HEK Media Panel covers a broad variety of human embryonic kidney 293 (HEK293) cell types but is not tied to specific transfection reagents or techniques. The panel comprises ready-to-use formulations designed to help AAV developers identify an optimal formulation for their target cell line. The media included in the panel are optimized for higher titers, are scalable and customizable, and come with process consultation and data analysis to expedite media optimization and current good manufacturing practice (CGMP) manufacturing.

The Applied Biosystems resDNASEQ Quantitative Plasmid DNA–Kanamycin Resistance Gene Kit is easy to use and offers an all-in-one solution for precise and rapid measurement of residual plasmid DNA. The kit includes a highly sensitive multiplex quantitative polymerase chain reaction assay and contains all reagents. It also offers optimized sample prep options to help streamline workflows and delivers results in under five hours. The kit targets conserved regions of multiple kanamycin-resistance gene families, covering all common plasmids with kanamycin resistant genes that are used in bioproduction for applications such as triple transfection of host cells to produce AAVs. According to Thermo Fisher in a company press release, residual plasmid DNA must be measured to demonstrate that it is below the thresholds set by regulatory agencies to ensure product quality and safety.

Thermo Scientific POROS CaptureSelect AAV affinity purification resins is the newest addition to the portfolio and includes POROS CaptureSelect AAV8, AAV9, and AAVX affinity purification resins as well as POROS CaptureSelect AAVX EvolveD Columns. These resins can significantly improve downstream processing of viral vectors by reducing purification steps. The resins also offer scalability to maximize process consistency, efficiency, and productivity, according to the press release.

"Despite the immense promise of gene therapy, robustness and yield continue to pose challenges in the manufacturing process," said Betty Woo, vice-president and general manager, cell and gene therapy at Thermo Fisher Scientific, in the press release. "Growth media that maximizes viral titer and rapid analytical tests contribute to cost-effective and efficient AAV production. Our expanding gene therapy portfolio is designed for a smooth ramp-up to clinical production, and we have made focused investments to ensure we stay ahead of commercial demand. Ultimately, our innovative solutions are developed to expedite the delivery of safer therapies to patients."

A new media panel, gene kit, and advanced resins that are now in Thermo Fisher Scientific’s gene therapy portfolio aim to reduce manufacturing costs and increase gene therapy viability.

Pharmaceutical Technology's In the Lab eNewsletter, August 2021