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查看更多产品信息 CTS™ Detachable Dynabeads™ CD3/CD28 - FAQs (A56996)
21 个常见问题解答
Superparamagnetic means that the Dynabeads magnetic beads exhibit magnetic properties when placed within a magnetic field, but have no residual magnetism when removed from the magnetic field.
This means that your targeted cells, proteins, or nucleic acids are only subjected to magnetic forces during the time the beads are on the magnet. The beads do not aggregate, but remain evenly dispersed in suspension.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
CTS Detachable DynabeadsCD3/CD28 (Cat. No. A56996) is shipped at 2-8 degrees C. We recommend storing it at 2-8 degrees C, protected from light.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Yes, CTS Detachable Dynabeads CD3/CD28 (Cat. No. A56996) and CTS Dynabeads CD3/CD28 (Cat. No. 40203D) preferentially isolate CD3+ CD28+ double-positive T cells, which are demonstrated to contain the clinically relevant subset of T cells (i.e., CD27 positive, PD-1 negative, CD8 positive CAR T cells).
Also note that CTS Dynabeads is the only isolation technology on the market that excludes exhausted T cells from the manufacturing of the T cell drugs. Please see this reference from University of Pennsylvania & Novartis that used CTS Dynabeads CD3/CD28 in a clinical study of patients with Chronic Lymphocytic Leukemia (CLL): https://pubmed.ncbi.nlm.nih.gov/29713085/
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Unlike magnetic beads from our competitors, CTS Dynabeads are not phagocytosed by T cells or incorporated into patients. CTS Dynabeads are inert to T cell phagocytosis and can therefore be removed (using the CTS DynaCellect Magnetic Separation System, Cat. No. A55867) before the final drug is injected into the patient.
There is also no evidence that CTS Dynabeads activate T cells non-specifically. Adding Dynabeads without antibodies, for example, to cell suspensions does not upregulate activation markers nor induces expansion.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
CTS Detachable Dynabeads CD3/CD28 (Cat. No. A56996) are coated with camelid-derived, single-domain (VHH) antibody fragments that identify and bind CD3 and CD28 markers, separately. These antibodies are produced in an Animal origin Free (AOF) yeast-based system, helping to reduce any risk of viral contamination.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
At the time of launch to market for CTS Detachable Dynabeads CD3/CD28 (Cat. No. A56996) in 2023, the shelf-life was 6 months, but our plan is to extend the shelf-life to 9, 12, 18, 24, 36, 48, and 60 months on a rolling basis. Therefore, our expectation is that the shelf-life will be 5 years by the year 2028.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
When using CTS Detachable Dynabeads CD3/CD28 (Cat. No. A56996) for simultaneous isolation and activation in one step, we recommend a 3:1 ratio of Dynabeads to target T cells. Please do not calculate it based on the number of total nucleated cells in your starting material.
When using CTS Detachable Dynabeads CD3/CD28 for activation only, downstream of a separate isolation step with a separate isolation reagent, the recommended ratio of Dynabeads to target T cells is 1:1.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Yes, CTS Dynabeads and CTS Detachable Dynabeads are GMP-grade and CTS-branded, which means that they are eligible for use in process development, clinical trials, and commercial manufacturing.
Thermo Fisher Scientific’s Cell Therapy Systems (CTS) translates to cGMP manufacturing, testing, and traceability documentation (such as Drug Master Files and Regulatory Support Files) for customer filing needs, and a proven track record of use in many clinical trials and commercialized drugs on the market.
The CTS Dynabeads portfolio, specifically, has been actively used in over 200 clinical trials in 2023 as well as in a number of FDA-approved drugs for commercial use.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
CTS Detachable Dynabeads are meant to be used with the CTS Detachable Dynabeads Release Buffer that comes in a bioprocess container compatible with our CTS DynaCellect Magnetic Separation System for automated workflows.
After the isolation step, the Dynabeads-bound T cells are incubated with the CTS Detachable Dynabeads Release Buffer to actively detach the cells from the Dynabeads. The CTS Detachable Dynabeads Release Buffer contains d-biotin that outcompetes a biotin derivative on the surface of CTS Detachable Dynabeads. It is not an enzymatic reaction.
Users may then rely on the CTS DynaCellect Magnetic Separation System to magnetically separate CTS Detachable Dynabeads from target T cells so they are left with a highly pure target T cell population in the positive fraction.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Yes, you would need to also purchase the CTS Detachable Dynabeads Release Buffer in order to take advantage of the active release mechanism for CTS Detachable Dynabeads CD3/CD28. The CTS Detachable Dynabeads Release Buffer contains d-biotin that outcompetes a biotin derivative on the surface of CTS Detachable Dynabeads CD3/CD28. It is not an enzymatic reaction.
After the active release of CTS Detachable Dynabeads CD3/CD28, you may use the CTS DynaCellect Magnetic Separation System to magnetically separate the CTS Detachable Dynabeads CD3/CD28 from target T cells so that you are left with a highly pure target T cell population in the positive fraction.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Due to the presence of biotin in the CTS Detachable Dynabeads Release Buffer, we do not recommend using CTS Detachable Dynabeads with media that already contain high amounts of biotin, such as RPMI media. Instead, we recommend pairing with media such as our CTS OpTmizer SFM and CTS OpTmizer Pro SFM.
Note: Internal studies have shown that the biotin in the CTS Detachable Dynabeads Release Buffer does not have any impact on cell phenotype or viability.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Dynabeads are not designed or intended to be used with centrifugation. Pressure from centrifugal forces should not theoretically be a problem for the Dynabeads themselves due to their rigid nature and composition. However, the force exerted by the Dynabeads on surrounding cells within the pellet may be detrimental to cell viability.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Each vial contains 4x10E8 Dynabeads per mL.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Thermo Fisher Scientific is the only supplier that has GMP-grade reagents with a technology that can both isolate and activate T cells in a one-step mechanism. This is enabled by CTS Detachable Dynabeads CD3/CD28 (Cat. No. A56996) where both CD3 and CD28 VHH antibody fragments are present on the surface of the CTS Detachable Dynabeads.
The advantage is that users do not need another reagent to perform activation steps downstream from target T cell isolation. Instead, they can save process time, steps, and additional costs that would normally be required had they used separate reagents for isolation and cell activation.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Dynabeads magnetic beads are uniform, non-porous, superparamagnetic, monodispersed and highly crosslinked polystyrene microspheres consisting of an even dispersion of magnetic material throughout the bead. The magnetic material within the Dynabeads magnetic beads consists of a mixture of maghemite (gamma-Fe2O3) and magnetite (Fe3O4). The iron content (Fe) of the beads is 20% by weight in our M-450 Dynabeads.
Dynabeads magnetic beads are also coated with a thin polystyrene shell which encases the magnetic material and prevents any leakage from the beads or trapping of ligands in the bead interior. The shell also protects the target from exposure to iron while providing a defined surface area for the adsorption or coupling of various molecules.
The uniformity of Dynabeads' size and shape provides consistent physical and chemical properties. These uniform physical characteristics lead to high-quality, reproducible results.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
This depends on the total number of cells that you are processing in a single run and the total number of Dynabeads used. However, our large-scale studies with 10-liter volumes on the CTS DynaCellect Magnetic Separation System yielded less than 25 residual Dynabeads per 3 million cells. And with 1-liter volumes, performance was even better as we observed less than 5 residual Dynabeads per 3 million cells (again in internal studies).
As a reference point, most regulatory agencies have an unofficial minimum threshold of 100 residual beads per 3 million cells.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
All of our CTS Detachable Dynabeads products are intended to be used with the CTS DynaCellect Magnetic Separation System. While small DynaMag magnets can be used to assess the functionality of the product in a laboratory scale, it does not allow you to see the full benefits or accurate performance should you decide to scale up to clinical or commercial manufacturing.
Using our CTS Detachable Dynabeads without the CTS DynaCellect Magnetic Separation System is not representative of the yield and purity obtained in the automated closed system.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Our CTS Dynabeads and CTS Detachable Dynabeads are supported with Drug Master Files (DMFs), Regulatory Support Files (RSFs), and other traceability documentation.
We have dedicated resources from our Quality and Regulatory teams to support customers who enter into a Commercial Supply Agreement with us.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
CD28- T cells in the starting material prior to manufacturing are associated with exhausted T cells and not relevant for the clinical efficacy nor persistence of the T cell drug in vivo in the patient.
Naive T cells require three essential signals for activation: T cell receptor (or CD3) engagement, co-stimulatory receptors (such as CD28) activation, and cytokine stimulation (IL-2 for expansion). Activated T cells downregulate the co-stimulatory molecule CD28 and loosen its expression altogether upon chronic antigen exposure. Downregulation of CD28 is a hallmark of senescent T cells. CD3+CD28- cells are therefore activated T cells and are susceptible to activation-induced cell death (AICD) if re-stimulated in the absence of the co-stimulatory receptor CD28. If isolated, these cells will die during the first hours of the expansion process.
Please see the following references:
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6600236/
https://onlinelibrary.wiley.com/doi/abs/10.1111/j.0105-2896.2005.00256.x
https://pubmed.ncbi.nlm.nih.gov/10882412/
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
Yes, CTS Detachable Dynabeads CD3/CD28 (Cat. No. A56996) and CTS Dynabeads CD3/CD28 (Cat. No. 40203D) preferentially isolate CD3+ CD28+ double-positive T cells, which are demonstrated to contain the clinically relevant subset of T cells (i.e., CD27 positive, PD-1 negative, CD8 positive CAR T cells).
Also note that CTS Dynabeads is the only isolation technology on the market that excludes exhausted T cells from the manufacturing of T cell drugs. Please see this reference from University of Pennsylvania & Novartis that used CTS Dynabeads CD3/CD28 in a clinical study of patients with Chronic Lymphocytic Leukemia (CLL): https://pubmed.ncbi.nlm.nih.gov/29713085/
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.
When used with the CTS Detachable Dynabeads Release Buffer, CTS Detachable Dynabeads CD3/CD28 (Cat. No. A56999) can be actively detached from target T cells at any desired time.
On the other hand, CTS Dynabeads CD3/CD28 (Cat. No. 40203D) that launched to market in 2006 may not be actively detached from target T cells. They passively dissociate from the target T cells post-isolation, over time, based on the binding kinetics of antibodies conjugated to the surface.
Please note that both products contain the same M-450 Dynabeads microspheres at their core but all other raw materials used, such as the antibodies, are different between the two products.
Find additional tips, troubleshooting help, and resources within our Dynabeads Cell Isolation and Expansion Support Center.