QuickGel™ technology is helping researchers overcome cell separation challenges in translational medicine, stem cell research and cancer therapy.
QuickGel™ technology is helping researchers overcome cell separation challenges in translational medicine, stem cell research and cancer therapy.
Quad Technologies has teamed up with Yu-Chang Bryan Wu, Ph.D. from the Randall Division of Cell & Molecular Biophysics at King’s College London, UK, to demonstrate the value of Quad’s MagCloudz™ Streptavidin Cell Separation kit for B-lymphocyte and antibody research.
Dr. Wu’s core research draws on parallel immune profiling of human B cells, which constantly diversify their immune repertoire by editing antibody-encoding genes in order to recognize the ever-evolving pathogens. His prior studies applied next generation sequencing and translational bioinformatics to probe changes in the B-cell repertoire for a better understanding of immunological mechanisms of ageing, vaccination and allergic diseases (https://kclpure.kcl.ac.uk/portal/yu-chang.wu.html).
At King’s College, Dr. Wu is currently working on an integrated approach to parallel profiling of B cells using multiple research systems. One of his major challenge is to enrich specific populations of B cells that can be stably transferred across different systems for streamlined analysis. Using conventional magnetic beads to isolate B cells, he has found experimental variations as a result of cell death induced by phagocytosed magnetic beads. MagCloudz Streptavidin purifies cells completely free of direct contact with magnetic beads. In contrast to conventional methods, the cells can remain immobilized on the gel “cloud” without the risk of internalizing magnetic beads after enrichment, when full release of the cells from the gel cloud is not required. Dr. Wu considered these options to represent a significant advantage of MagCloudz Streptavidin platform over conventional magnetic bead enrichment kits and was excited to test the MagCloudz Streptavidin platform to isolate B cells in his research projects.
In a pilot experiment, Dr. Wu tested the feasibility of isolating human primary B cells from cryopreserved PBMC samples, using the MagCloudz Streptavidin platform in combination with biotinylated anti-CD19 and anti-CD20 antibodies. Following MagCloudz enrichment procedures, cells were released from the gel clouds, characterized by multi-color flow cytometry, and compared to unenriched samples.
Left: The initial PBMC sample contained 9.35% CD3-/CD19+ B cells, which were targeted with biotinylated anti-CD19 and anti-CD20 antibodies and added to MagCloudz™ Streptavidin.
Right: The post-enrichment sample contained 98% of CD3-/CD19+ B cells.
MagCloudz™ enriched B cells (right) contained multiple cell subpopulations at similar frequencies as those in the unenriched sample (left). Q1: Classical memory, Q2: IgM memory; Q3: Naïve; Q4: Double negative memory. Other markers, including IgA, IgG, IgM, IgE, CD38 and CD138, were present at comparable values between two setups.
The viability of B cells in cryopreserved PBMC samples were assessed by Live/Dead Aqua staining.
(Left) 91.3% of B cells were viable in unenriched samples. (Right) 90.6% of enriched B cells were viable.
These data indicate that MagCloudz Streptavidin can isolate B cells of high purity and viability without altering their phenotype during the enrichment procedure. Dr. Wu concluded that: “We are very encouraged by the capability of the MagCloudz platform to obtain B cells with great efficiency, and impressed by its distinct feature to immobilize cells on the solid phase of gel clouds without killing these cells. These immobilized B cells are now put through a stream of experiments in different research systems. We expect to integrate MagCloudz into current workflow and explore its potential in other area of our projects.”
Nithya Jesuraj, Ph.D
When given the opportunity to join the team at Quad Technologies as a Research Scientist, I was excited because I realized that the QuickGel technology platform had great potential to improve cellular bioprocessing methods critical to advancing cancer treatments and address challenges faced by researchers in this field.
In research during my graduate tenure at Washington University in St. Louis, my work focused on developing transplantation therapies for the treatment of peripheral nerve injuries. In this promising cell therapy application, the biggest challenge I faced related to cell purity within transplants, namely the contamination of therapeutic Schwann cells with unwanted cell types such as fibroblasts. Conventional cell purification approaches did not provide a means to solve this cell purity challenge.
After joining Quad Technologies from Boston Biomedical, where I spent 3 years designing and developing RNAi therapeutics to treat cancer, I was excited to continue contributing to the advancement of revolutionary cancer treatments. One of the first projects I started working on at Quad Technologies was developing a method for activation and expansion of T Cells using the MagCloudz cell isolation kit. Taking advantage of its compatibility for use with any antibody, I was able to demonstrate that MagCloudz provides a platform for uses beyond cell isolation. Specifically, we have shown that MagCloudz-based T cell expansion results in population doublings comparable, and in some cases better than existing competitors, with the added valuable benefit of our simple magnetic particle release mechanism.
I am very excited about the promise of recent data showing that MagCloudz can activate and expand T Cells while preferentially enriching for CD8+ cells expressing critical memory and naïve phenotypic markers. The presentation of this data at the American Society of Hematology (ASH) meeting this month is a great accomplishment for the Quad team, and I’m very proud to have played an integral role. Additionally, this project has provided me the opportunity to work collaboratively with external researchers and experts in the T Cell field to refine ideas and experiments based on specific needs of the clinical bioprocess workflow for autologous immunotherapies. I continue to be motivated as I uncover new questions and challenges to address head on!
As Quad continues to grow and expand its portfolio of QuickGel technology based products for cell therapy applications, I look forward to learning more from our customers, tackling technical challenges, and contributing to the advancement of solutions that can help defeat cancer and other incurable diseases.
Type 1 diabetes (T1D) is thought to develop from an imbalance in a population of white blood cells referred to as T helper cells. In T1D, a population of destructive T cell recognizes the insulin producing pancreatic beta cells as foreign and causes beta cell destruction leading to high blood sugar levels. In individuals without autoimmune diseases like T1D, a unique population of regulatory T cells (Tregs) keeps these destructive T cells in check.
The Todd M. Brusko Laboratory is developing clinical protocols to isolate and expand Tregs isolated from a patient’s own supply of cryopreserved umbilical cord blood (UCB). The hope is that this newly isolated population of Tregs from UCB will create a unique cellular therapy that will aid in the restoration of immune tolerance to pancreatic beta cells. We believe this state of immune tolerance to be an essential component to any effective therapy, whether it be early prevention, disease reversal at onset, or restorative therapies following transplantation. We are currently testing MagCloudz Streptavidin as a tool for clinical trials to help shorten the time it requires to purify CD4+CD25+CD127lo/- Treg from UCB by florescence activated cell sorting (FACS). Using CD4-positive selection with releasable MagCloudz would shorten the isolation procedure by hours without interfering with post-isolation in vitro stimulation, as well as decrease CD8+ effector T cell contamination that can cause the final adoptive cell therapy product to fall short of the clinical release criteria.
Additionally, we are optimizing double positive-selection techniques to aid in the isolation of CD4+CD25+CD127lo/-Treg from human organs, were these cells are extremely infrequent. Using samples provided by the Network for Pancreatic Organ Donors with Diabetes (nPOD, jdrfnpod.org), we are studying the adaptive immune cell receptor repertoire in T1D. Prior studies characterizing the T cell and B cell receptor repertoires in T1D have been restricted to peripheral blood due to inaccessibility of the spleen or pancreatic-draining lymph nodes (pLN) and the inherent dangers associated with pancreatic biopsy. In an effort to determine whether such studies broadly reflect the distribution of T and B cells present in the target organ and tissue-associated lymph nodes in T1D, we purify Treg, Tconv, CD8+ T cells, and B cells by FACS from the pLN, spleen, non-pLN, peripheral blood, and pancreatic islets of transplant organ donors obtained by the nPOD program. We are currently testing the feasibility of using MagCloudz in a unique double positive-selection technique to enrich for Treg and shorten the time for isolation by FACS from 12 hours to less than one.
In summary, we are very excited about our relationship with Quad Technologies. From our experiences, the addition of MagCloudz to our toolbox for immune studies help overcome one of the main limitations of isolating CD4+CD25+CD127lo/- Treg by FACS without sacrificing purity.
Alaina Peterson, M.S.
Field Applications Specialist, Quad Technologies
Joining the Quad Technologies team as the Field Applications Specialist has been such an exciting experience. Prior to joining the dedicated Quad team, I have spent 5+ years working in various settings in the biotech and pharmaceutical field. Specifically, I worked as research scientist in various immunology related fields including cellular immunotherapy, immuno-oncology and allergies.
I am looking forward to bridging the gap between Research and Development and technical customer support at Quad Technologies. While I enjoy running experiments, I am eager to step away from the lab bench at times and have more interface with customers. I quickly realized in my past work that it takes a specific individual with a specific skill set to translate what is happening in the lab into layman’s terms that the end user is able to fully understand. Research is not successful if the end user cannot understand the protocol, application or technical note. I am eager to work with MagCloudz™ and Quickgel™ technology in the lab here at Quad, and strive to talk about my findings in big pharma or academia settings. Coming off the research bench, I am really excited to interact with other scientist and to help optimize the MagCloudz™ assay for their individual and specific needs.
My hopes are to see MagCloudz™ used for targeted cell therapy as well as the technology used in clinical trials. I have gained a strong belief that immunotherapy will revolutionize medicine for every patient, whether they have Alzheimer’s or cancer. I believe the MagCloudz™ technology will provide critical support for the immunotherapy field: one of sciences’ fastest growing areas of research.
Research and Development Scientist, Quad Technologies
When I joined Quad Technologies in the spring of 2014 the company was still in its infancy and as its third employee, I made an immediate impact as we strived to make the MagCloudz™ cell separation kit a reality. United behind the idea of enhancing cell separation workflows within the life sciences industry, the small Quad team endeavored singularly to deliver an exceptional customer solution, by the way of a magnetic-label-free alternative to conventional cell separation strategies. Our impetus was QuickGel™, the hydrogel polymer that would go on to become the platform for the MagCloudz cell separation kit and revolutionize cell separation by providing customers with happy, magnetic-label-free cells.
The challenge of our day-to-day work was finding a way to take MagCloudz potential on paper and translate it into a tangible product.
Specifically, to meet the needs of our customers we revamped, fine-tuned, and updated chemistries, which in turn led to increased cell yields and purities for the end-user. We implemented stringent methods development, working closely with external collaborators to institute effective protocols which led to consistent, reliable results. In addition, we efficiently scaled our manufacturing processes reducing our own costs and moving the savings onto the research and industry institutions utilizing our technology. These improvements and developments have allowed us to take the MagCloudz cell separation kit from our own laboratory and place it in the hands of capable researchers on a global scale.
Having been instrumental in the development of Quad Technologies’ first product, it is my hope to contribute towards subsequent products. With our know-how and technology portfolio Quad Technologies is in a unique position to tackle the challenges of the health and life sciences industries, specifically by increasing the affordability and scope of personalized medicines. When some cancer drugs are only effective in 25% of patients, there is a distinct need for customizable, targeted cell therapies and drug delivery vehicles.
As someone on the ground floor I have had the experience of watching a start-up become a small company and seeing the number of employees triple over the past two years. I have also seen what it takes to bring a product from development to the market place. I know that we will continue to grow and that we will continue to offer solutions in the cell separation space and beyond. I can’t wait to see what is next!
Andrew Ball, Ph.D.
Vice President of Research and Development, Quad Technologies
I work as VP of Research and Development at Quad Technologies and strongly believe that there has never been a better time to be involved in cell therapy research. Many research breakthroughs of the past 20 years are proving to have real translational value and a new generation of cell therapies are successfully entering the clinic. The next great challenge is delivering cost-effective scale-up of cell therapies. We’re moving from asking ‘is cell therapy possible?’ to ‘how can we make cell therapies more robust, accessible, and affordable?’ – this latter question is driving everything we do at Quad Technologies.
At Quad my responsibilities include overseeing research, development, and manufacturing operations. We recently commercialized our first product, the MagCloudz™ Streptavidin cell separation kit, built around a dissolvable hydrogel platform, QuickGel™ which represents a disruptive technology for cell separation. My ongoing focus is to help Quad develop a strong portfolio of innovative technologies and products which will ultimately improve both the efficacy and safety of cell therapies, while simultaneously increasing manufacturing efficiency. It’s a huge – but a hugely exciting – challenge.
Prior to joining Quad, I worked in both academic and industrial settings, seeking in both to address the ongoing need for improved cellular models and assays to understand the pathophysiology and pharmacology of disease, and to develop cellular models which could be engineered and efficiently scaled to treat human diseases. At Quad, my work has focused on the opportunities and challenges associated with clinical applications of cell transplantation, particularly in the emerging field of T cell-based cancer immunotherapy. The enormous promise of this field is tempered by significant manufacturing challenges associated with phenotypic reproducibility and, particularly, cost of goods, and also by unwanted, often life-threatening side effects associated with cell therapies.
A key challenge that remains is to make cell therapies a reality for the millions of patients worldwide with diseases like cancer and diabetes. We know that cures are right around the corner, and in some cases, already here – but the dream is to make these accessible to many more patients. At Quad we have a great team and a great technology, and are ready to play a significant role in the cell therapy-driven transformation of healthcare which is upon us.
Michael Brehm, Ph.D.
Program in Molecular Medicine
University of Massachusetts Medical School
Quad Technologies’ has partnered with Michael Brehm, Ph.D and Rita Bortell, Ph.D from the University of Massachusetts Medical School Program in Molecular Medicine to evaluate the performance of MagCloudz™ Streptavidin in their research applications. Brehm and Bortell’s research focuses on the development of new and improved therapeutic measures for patients suffering from Type-1 diabetes (T1D). T1D is an auto-immune disorder, which occurs when ‘germ and virus fighting’ T-Cells attack the body’s own organs and tissues instead of foreign pathogens. In T1D, these renegade T-Cells cause destruction of insulin-producing pancreatic beta cells, effectively preventing insulin production and causing the T1D disease state.
At UMass, Brehm and Bortel are working on ways to study how auto-immune diseases, such as T1D, progress and develop new therapeutic approaches using ‘humanized’ mouse models. These special ‘humanized’ mice closely mimic a normal human immune system and therefore provide an excellent platform for disease research. However, the major challenge with using humanized mice is that despite all the time and effort involved, not every mouse successfully develops a ‘human’ immune system.
Current protocols call for direct transplantation of human T-Cells, isolated from fresh blood samples, into mice which lack an immune system of their own (severely compromised immunodeficient/SCID mice). The human T-Cells essentially ‘take over’, thereby creating a fully functional mouse with a ‘humanized’ immune system. According to Brehm and Bortell, their major challenge is that conventional methods for isolating the T-Cells from blood using magnetic beads are expensive, slow, laborious and at the end of the process, the T-Cells are coated in magnetic beads. The presence of these beads on the cell surface reduces the T-Cells’ ability to proliferate and causes cell death, which greatly reduces the efficacy of the transplantation and engraftment process in mice.
For this reason, Brehm and Bortell were very excited to test the MagCloudz™ Streptavidin Cell Separation Kit in their labs. The MagCloudz cell separation kit has significant advantages over conventional magnetic bead isolation approaches because our QuickGel™ technology enables the T-Cells to be fully released from the magnetic bead carriers. This unique release capability improves long-term cell viability and should theoretically improve the transplantation and engraftment rate of these isolated T-Cells in SCID mice.
We tested this hypothesis in a feasibility study, where conventional magnetic bead T-Cell isolation methods were compared to our MagCloudz™ Streptavidin platform using a biotinylated CD3 antibody to target the desired T-Cells in the initial blood sample. T-Cell uptake, purity and viability at the end of the separation process was compared directly between the two methods, shown in the figures below:
The initial blood sample contained 12.1% CD3+/CD45+ T-Cells, which were targeted with a biotinylated anti-CD3 antibody and added to MagCloudz™ Streptavidin or conventional streptavidin magnetic beads.
Excellent T-Cell uptake from the initial blood sample was demonstrated in both approaches.
High purity of the recovered T-Cell populations: 95.9% with the MagCloudz™ Streptavidin and 95.6% with the conventional streptavidin magnetic beads.
Immediately following the separation protocol, the viability of the recovered T-Cells was determined using annexin-V (an apoptosis marker) and 7-AAD (a membrane permeability marker). Viability was >90% for both approaches, with the MagCloudz™ Streptavidin kit showing higher viability at 94.5% than the conventional approach.
These data indicated that MagCloudz™ Streptavidin perform at or above the level of the conventional magnetic separation approaches for isolating T-Cells from whole blood. Brehm summarized that: “Our goal is to have high purity, highly viable T-Cells that are able to proliferate and effectively generate a humanized mouse. The potential for scaling-up using the MagCloudz platform to process large numbers of blood cells and transplant larger groups of mice than conventional methods allow is very exciting for us.”
Further studies which examine long-term cell viability and proliferation are underway, but we anticipate that T-Cells isolated using the MagCloudz™ Streptavidin will have improved transplantation and engraftment rates in humanized mice.