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Successful process development minimizes cost while enhancing the efficiency and quality of T Cell and NK Cell therapies1. R&D Systems' immunoassay solutions expedite immune cell therapy development and manufacturing workflows. Select from the most flexible R&D Systems™ Luminex® assays*, industry-leading Quantikine™ ELISA kits, or Simple Plex™ Assays on the Ella™ instrument, a rapid and hands-free precision biomarker detection instrument.
Table of Contents
Quantify GMP and Secreted Proteins
Immunoassays are required to confirm the precise addition and clearance of GMP growth factors during the cell culture phases of the immune cell therapy workflow. Excessive growth factor exposure can cause T cell overactivation and exhaustion. Quantikine ELISA kits, automated Simple Plex assays on the Ella instrument, and Luminex assays can be used to quantify GMP and secreted cytokines during the cell culture, activation, and expansion stages of the workflow.
GMP-grade human IL-2, IL-7, and IL-15 (green circles) were interpolated with the appropriate Quantikine High Sensitivity ELISA Kit standard curve (blue circles) using 4PL logistic regression analysis.
GMP-grade human IL-2, IL-7, and IL-15 (green circles) were interpolated with the appropriate standard curve (blue circles) using 4PL logistic regression analysis. Automated Simple Plex assays on the Ella instrument return data in 90 minutes or less.
Assess T Cell Activation
Immune cell therapy efficacy is influenced by a number of factors including T cell activation, which must be evaluated during the manufacturing workflow. Key T cell activation markers include Granzyme B, IFN-gamma, IL-2, Perforin, and TNF- alpha. Automated T cell activation marker assays on the Ella instrument include a built-in factory standard curve, up to 4 logs of dynamic range, and 21 CFR Part 11 compliance for cell therapy manufacturing processes. These assays are vertically integrated with R&D Systems antibodies to deliver reproducible data you can trust. Quantikine ELISA kits, the most referenced ELISA on the market are also available to profile the T cell response.
Download the Cell Killing Application Note
T cell activation markers are robustly elevated when CD19 CAR T cells are co-cultured with CD19+ Nalm-6 cancer cells, but not CD19- K562 cells. T cell activation markers Granzyme B, TNF-α, IFN-γ, and IL-2 were assayed in parallel using a 32x4 Multianalyte cartridge on the Ella instrument. Bar graphs: mean ± SD (n=3).
Automated Viral Titer Assays
Viral vectors are a popular tool for gene transfer in immune cell therapy workflows. Control of critical quality attributes (CQAs) such as viral titer is vital for maintaining therapeutic consistency. By automating the AAV and Lentiviral titer quantification process, Simple Plex viral titer assays on Ella decrease the time to result and reduce user error thereby enhancing team productivity and product consistency. Our viral titer assays are developed in collaboration with AAV industry leader PROGEN.
Purchase Assays: AAV1 Viral Titer Assay | AAV2 Viral Titer Assay | AAV6 Viral Titer Assay | AAV8 Viral Titer Assay | HIV-1 p24 Viral Titer Assay
Linear correlations of the Simple Plex AAV viral titer assays and PROGEN AAV ELISAs. The aggregated Simple Plex and PROGEN ELISA data were plotted and a linear fit was applied to the data for each AAV serotype. The Simple Plex AAV viral titer assays are correlated with the corresponding PROGEN ELISAs as follows: AAV1 R2=0.99, AAV2 R2=0.99, AAV6 R2=0.99, AAV8 R2=0.99. The data showed a nearly perfect correlation where the quantitative range of both viral titer assays overlap.
The linearity of dilution was evaluated using in-process samples across multiple matrices. Each in-process sample showed good dilutional linearity at all 4 dilutions tested, with 80-120% percent recoveries compared to the minimum required dilution (MRD).
Automated Process Impurity Testing: HEK 293 HCP | Endonuclease
Human Embryonic Kidney (HEK) 293 cells are used for the production and scale-up of viral vectors. HEK host cell proteins (HCPs) must be cleared via in-process testing to avoid adverse host responses. Ella's automated HEK HCP process residual assay is developed in partnership with Cygnus Technologies. This assay recognizes thousands of HCP species, satisfies the demands of timely data generation for maintaining efficient production, and overcomes the lengthy investigations and sample retests that often result from human error.
Purchase the HEK 293 HCP Assay
The Simple Plex HEK 293 HCP 3G assay is highly correlated (R²=0.9799) with the Cygnus HEK 293 HCP ELISA Kit, 3G Resupply 1.
Detection of HEK HCPs during bioproduction with the Simple Plex HEK 293 HCP 3G assay and the Cygnus HEK 293 HCP ELISA Kit,3G Resupply 1 ELISA. Three industrial purification processes were profiled using the Cygnus ELISA (green), and the Simple Plex assay (blue). These processes were measured at multiple steps (X-axis) to show the elimination of HEK HCP from the product. Across each process, the Simple Plex assay demonstrates the same fold reduction in HEK HCP as the Cygnus ELISA although some differences in absolute sample values were observed.
Rapid Endonuclease Quantification
Endonuclease is useful for applications where host cell nucleic acid digestion is necessary. Such applications include viral vector preparations, vaccine products, and other biotherapeutics. Once the host-cell nucleic acid is digested, one must verify that residual endonuclease has been cleared from the final product. The Simple Plex Endonuclease assay is an easy-to-use immunoassay for detecting residual endonuclease impurities in bioprocess samples.
In Vivo Monitoring
R&D Systems Luminex Assays
Cytokine release syndrome (CRS) is an adverse response to CAR T cell therapy. IFN-γ, IL-6, IL-10, and TNF-alpha are the core cytokines involved in CRS. R&D Systems Luminex High Performance Assay Panels are validated for use with human serum and plasma samples. Select from a premixed, user-mixed, or fixed panel, based on your needs.
The XL Cytokine Performance Panel detects the inflammatory response of CRS biomarkers. These graphs visualize the quantification of CRS analytes in Peripheral Blood Mononuclear Cell (PBMC) supernatants before and after stimulation with anti-CD3 epsilon and anti-CD28.
Purchase Preclinical & GMP Proteins and Immunoassays
| Target | Preclinical Protein | GMP Protein | Quantikine ELISA | Simple Plex Assay | Luminex Assay |
| Granzyme B | Human | yes | yes | yes | |
| IFN-gamma | Human | yes | yes | yes | yes |
| IL-1 beta | Human | yes | yes | yes | yes |
| IL-2 | Human | yes | yes | yes | yes |
| IL-4 | Human | yes | yes | yes | yes |
| IL-6 | Human | yes | yes | yes | yes |
| IL-7 | Human | yes | yes | yes | yes |
| IL-8 | Human | yes | yes | yes | |
| IL-12 | Human | yes | yes | yes | |
| IL-15 | Human | yes | yes | yes | yes |
| IL-18 | Human | yes | yes | yes | |
| IL-21 | Human | yes | yes | yes | |
| IL-23 | Human | yes | yes | ||
| IL-27 | Human | yes | yes | ||
| Perforin | yes | ||||
| TGF-beta 1 | Human | yes | yes | yes | |
| TNF-alpha | Human | yes | yes | yes | yes |
This list is not exhaustive. Don't see what you need? Search for your target using the search bar at the top of the page.
Bulk Immunoassay Services
Did you know that Bio-Techne is vertically integrated? Vertical integration enables us to control our supply chain and enable a consistent cell and gene therapy development and manufacturing workflow. Bio-Techne is a partner that you can grow with.
Need help with an immunoassay-specific project? Start a conversation with an Immunoassay Specialist.
Bulk ELISA Assay Services | Bulk Luminex Assay Services | Bulk Ella Services
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Campbell, A et al. (2015) Concise Review: Process Development Considerations for Cell Therapy Stem Cells Translational Medicine 4:1155. PMID: 26315572.
*Luminex® is a registered trademark of the Luminex Corporation - A DiaSorin Company
