On-Site Quality Control
Efficient on-site quality control of cell-based products
In this century, development of Advanced Therapy Medicinal Products (ATMP), including gene therapeutics and human cell therapeutics, has been envisioned as ‘the’ beneficial breakthrough, capable of significantly advancing human health in the near future.
This innovative medical approach bears the potential of curing a braod spetrum of human diseases, ranging from cancer to heart disease and immunological disorders.
Cell therapy manufacturing processes comprise consistent procedures of culture, expansion, genetic manipulation, stimulation, and in case of treatments of multiple patients, cell banking. Therefore, manufacturing procedures as well as analytics of human cell therapeutics need to be adapted to this broad range of complex manufacturing processes ensuring adequate safety, purity and potency of the final products.
You are looking for a method to monitor each step of production and to control quality of your cell-based products, but ...
BioRam® is an all-in-one device that allows reliable screening of each step during production and efficient validation of the completed cell-based products.
Raman spectra provide even more detailed information about intermediates and finalized therapeutics as compared to antibody dependent FACS analysis or immune fluorescence. For quality survey with BioRam® no labels or antibodies are required and only a few 100 cells are sufficient. Taken altogether BioRam® warrants fast amortization.
BioRam® saves time and money with each quality assurance measurement!
Examples:
1. BioRam® saves costs and shortens hospitalization:
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a: Quality assurance of autologous skin grafts
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b: Survey of successful transfection with anti VEGF-vector to treat exudative macula degenerata (wet-form)
2. BioRam® saves life and money:
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a: Quality control of blood products prior to transplantation
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b: “Fitness” test of frozen cells after thawing
3. Raman Spectroscopy:
A noninvasive analysis tool for the discrimination of human skin cells
4. Raman spectroscopy as a tool for quality and sterility analysis for tissue engineering applications like cartilage transplants
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you can‘t get the right antibody?
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you don‘t want to loose too much valuable material?
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you want to reduce quality assurance costs?
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you need immediate results for your decisions?
1. BioRam® saves costs and shortens hospitalization:
a: Validation of patient derived skin graft
Quality control of cell-based products is essential to ensure safety and efficacy of the product before using it for patients. Deep skin wounds heal badly, developing into debilitating, permanent, disfiguring scars that require intense medical and home care. The project EuroSkinGraft deals with a novel generation of skin substitutes to clinically treat a broad spectrum of severe skin defects. The first bioengineered, autologous, dermo-epidermal skin grafts are presently undergoing clinical trials.
A 2x2cm piece of skin is taken from the patient. Cells such as keratinocytes, melanocytes, fibroblasts and endothelial cells are isolated, proliferated and cultivated on biodegradable matrices. The unambiguous identification of cells and their counting after collection from a donor is difficult. As the different cell types have different cell division rates, the final tissue composition is difficult to determine. At the moment FACS analysis is performed during production to discriminate cell population and to count their number of cells applied onto the skin graft. DNA count out of a piece of the ready-to-be-applied skin graft is used to check number of overall cells. None of the technologies can be applied without affecting the cell viability or to give insight into the functionality or quality of the cells within the final product.
BioRam® performed successful measurements of expanded cells (from healthy skin biopsies) and provided quality tests of the final graft. This showed that Raman spectroscopy has clear advantages over conventional methods:
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No labeling with markers, colors, antibodies or beads needed
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No modification of cell surfaces or activity state
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No physical stress through shear forces or non-physiological conditions
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No destruction of biological material
In summary about 3.000 Eur could be spared including time for expensive cell expansion with corresponding culture media and agents as well as costly antibodies plus valuable patient time in intensive care.
CellTool is currently in the process to achieve GMP validation from Swissmedic for this autologous skin graft.
b. Survey of successful transfection
The EU consortium TargetAMD (www.targetamd.eu/) aims on treatment of age-related macula degeneration (AMD) which is a major cause of blindness in elderly people in industrialized countries. There exists the atrophic (“dry”) and the exudative (“wet”) form, the latter being the most devastating one. The exudative form of AMD is characterized by the presence of uncontrolled vascularization, driven by the angiogenic vascular endothelial growth factor (VEGF).
Currently exudative AMD is treated with frequent intravitreal injections of the VEGF inhibitors such as pegaptanib, ranibizumab, and bevacizumab resulting in significant visual acuity gains in 30% to 40% of patients, while arresting the disease development in 90% of the remaining patients. However, anti-VEGF therapy is cost-intensive, and is accompanied by significant side effects, such as submacular hemorrhage and possible arterial thromboembolic events.
Thus, a gene therapeutic approach by delivering the PEDF gene directly to retinal cells should overcome those complications. To ensure an efficient delivery of the PEDF gene and its integration into the host cell’s genome, electroporation to support the plasmid-based integrating Sleeping Beauty transposon system (SB100X) is used which was established recently as a safer alternative to integrating viral vectors.
With BioRam we could monitor the transfection rate within less than an hour checking about 100 cells. Depending on the patient transfection rate was between 15% and 30 %.
At present BioRam® is the only technology that can monitor efficacy of transfection within the required time between extraction of the retinal cells and re-inoculation of the treated cells – especially as in this case cells are rare.
CellTool will participate at the phase II clinical trial to validate BioRam® as quality standard for the advanced treatment of exudative AMD.
2. BioRam® saves life and money:
a: Quality control of blood products
Even today, guaranteeing the quality of all blood products is difficult to achieve. Only about 1% of the blood bags are actually tested for quality and purity during the production. Sofar there is no control of blood products immediately prior to transfusion. The combination of Raman spectroscopy with optical trapping provides a non-invasive, fast and easy approach for ensuring quality of blood products directly before use. This on one hand enhances patient safety but also saves money as blood products can be tested individually and do not need to be discarded due to empirically determined dates of expiry.
b: "Fitness"test of frozen cells after thawing
Most of cell based therapeutics are froozen for storage. Well defined freezing protocols exist for the different types of AMTPs (advanced medical therapeutic product). However, for thawing are not yet standard procedures available.
BioRam® could depict differences between freezing time and time post thawing in a fast and easy manner.
Thus, protocols might become established to warrant best quality and functionality of the AMTP for patient safety and efficacy of the treatment.
