How lyophilization can open doors to new results and even help promote equity in studies
We’ve made great strides in global disease research, advancing discovery and improving our understanding of disease biology. However, some populations remain underserved: Logistical challenges in sample preparation and reagent stability often mean remote and rural communities around the world are underrepresented in studies, which can contribute to health inequities. Implementing unique, out-of-the-box workflows and technologies to enable the collection of high-quality data is therefore key to promoting research equity.
Enter lyophilization. Also known as freeze drying, lyophilization is a dehydrating process in which water is removed from previously frozen samples by sublimation (the transition of water directly from ice to vapor), maintaining the chemical structure of the original product. Lyophilizing an assay prevents degradation of components such as enzymes, antibodies, proteins, DNA or oligonucleotides, resulting in advantages like:
- Enhanced stability
- Easier transportation
- Long shelf life
The applications of lyophilization are diverse: It’s used in the food industry (for example, for astronaut meals or instant coffee), archeological conservancy (drying organic artifacts), archival conservation (saving water-damaged documents or books) and even taxidermy (long-term bacteria-free preservation).
In the pharmaceutical field, lyophilization is a key process for the preservation of biopharmaceuticals. It’s often used to help stabilize active pharmaceutical ingredients (APIs) and compositions that are unstable in other forms, and since the drying process doesn’t require heat, it’s well suited for sensitive biologics and APIs. Furthermore, lyophilization is also used to increase solubility and bioavailability of poorly soluble drugs, making it an important tool for maintaining the efficacy and safety of pharmaceutical products and a reliable choice for emergency supplies.
Thanks to their ability to support advanced drug delivery and streamline clinical operations, lyophilized products are a common choice for research. From monoclonal antibodies to vaccines, lyophilization is a key step in helping develop immunoengineering technologies being investigated in clinical trials. It’s also been used in such trials as studying the effects of fecal microbiota transplantation, looking to repair acne scars, developing a method for corneal transplants, observing the use of growth factors in knee osteoarthritis and even assessing anaerobic performance in football players, to name just a few, proving that the uses of lyophilization in clinical research are diverse and well established.
Groundbreaking study successfully includes remote locations using lyophilized panels
Researchers from the University of Sydney, led by Helen McGuire, PhD, looked to use the power of lyophilization to address the challenges of including resource-limited and rural sites in immune system studies. The group had already used a comprehensive 38-parameter immunophenotyping CyTOF™ panel to develop a blood-based immune signature that reported to be associated with differences in response to checkpoint therapies targeting the PD-1/PD-L1 pathway in melanoma and lung cancer. Wanting to apply these findings in remote settings with an expanded panel, they turned once again to Standard BioTools™ technology.
The researchers found that recent developments in CyTOF immune monitoring provided a simplified workflow for asynchronous sample collection. McGuire and team used a LyoMax CyTOF Panel because it could be implemented without specialized training or tools, harnessing the power of “cutting-edge data made possible without cutting-edge equipment.” The stability of heavy metal-conjugated antibodies and the lack of signal from autofluorescence meant stained samples could be cryopreserved and stored long-term, and the dried-down cocktail was deemed “simple to use,” yielding reproducible staining with little required sample.
McGuire’s team implemented their workflow across Australia, with 14 cancer patients recruited at three sites over four months. Remote asynchronous sample collection and surface staining was performed. Samples were then shipped to a central lab for multiplexed barcoding, which minimizes technical variation, and intracellular staining of several key functional markers. Analysis was performed in CellEngine, and principal component analysis of all samples demonstrated harmony across the sites. Overall, the team concluded that the large size of the panel and the ultimate flexibility of CyTOF technology resulted in a simple yet robust workflow for multi-site clinical trials. Furthermore, they found that the “workflow enables access to underserved communities, facilitating equity in immune phenotyping studies.”
Going forward, this research could have practical applications for improving study design and research stratification approaches in underserved communities, ultimately improving the equity and scalability of immunophenotyping studies to better serve people who live in geographically dispersed areas.
The unique benefits of mass cytometry ensure steady, dependable performance and improve operational burden, from labs to remote hospitals. CyTOF assays have been shown to be stable across experimental conditions and eliminate the need for additional controls and titrations, reducing technical variation and improving reproducibility.
In addition to the validated dry-format Maxpar™ Direct Immune Profiling Assay, custom lyophilized LyoMax™ Panels used in this study provide the same streamlined workflow but are tailored to specific markers of interest and standardized for large batches. LyoMax Panels are built to support large-scale, longitudinal and multi-site studies and can be customized to program specifications, ultimately providing deep insight into disease biology and immunophenotyping research through comprehensive immunophenotyping across a range of sites, regardless of logistical challenges.