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Next Generation Solutions for Immune and CAR T Cell Therapy

The next generation of immune cell therapies require a new generation of analysis tools
A safe,?potent, and persistent immune cell product depends on developers mustering the full repertoire of immune cell function; activation,?proliferation, cell fate, cytotoxic killing, immunomodulation, and memory. All of which must be accomplished in the ever-changing, suppressive, and toxic tumor microenvironment.
Agilent is dedicated to supporting these next-generation therapies, providing key technologies to enable high-efficiency gene editing and assessment of real-time cell function, phenotype, and fate. Used together, this tool bench provides the capability to measure and control immune cell function. Thus enabling translational researchers and developers to achieve the necessary level of therapeutic potency and safety.
Carl June
Breakthroughs in Immunotherapy
- A chat with Carl June
David Ferrick from Agilent interviews Carl June, renowned Immunologist and immunotherapy pioneer.
?Measuring phenotype, potency, and persistence at all phases of development
The Agilent immunotherapy tool bench covers the iterative cycles investigators need in early discovery. Stable, on target CRISPR editing combined with immunophenotyping (NovoCyte/Quanteon) and the ability to assess and validate edits on live-cell platforms that deliver relevant, time-resolved functional data (xCELLigence/Seahorse XF).
Test gene edits and other interventional strategies for phenotype, killing efficiencies, persistence, on/off target effects, and cell fate programming. Then relate this data to inhibitory effects such as exhaustion, anergy, and other escape phenotypes due to interactions with a constantly evolving tumor and its microenvironment. Agilent xCELLigence and Seahorse XF analysis can sensitively measure live cell function and simultaneously scan the temporal components of an ever-evolving immune response in terms of efficacy, persistence, and safety.
Agilent Seahorse XF and xCELLigence platforms are being evaluated by various development and production groups to define key quality and process performance criteria impacting potency and efficacy. As processes move from development to production, seamlessly transition to SureGuide guide RNAs manufactured under GMP using the same synthesis technology.
Discover the Agilent immune cell therapy tool bench
xCELLigence Real Time Cell Analysis (RTCA)
Quantify dynamic cell behavior that can be missed by labor-intensive endpoint methods to gain a more comprehensive measure of cancer cell killing including potential escape phenotypes.
xCELLigence RTCA technology provides a label-free, robust, and simple workflow, enabling measurement of diverse effector cells and molecules for cytotoxicity. It is a platform for more sensitive in vitro potency assessment that helps mimic activities in vivo.
Percentage cytolysis of target PC3 (Prostate) cancer cells by NK92 effector cells are measured on the xCELLigence system, starting at effector to target ratios as low as 0.625:1.
Agilent Seahorse XF analysis
Discover and tune immune cell programming to mount a sustained and robust antitumor response. Real-time metabolic analysis enables you to measures and tune the drivers of immune cell fate and function. Quantify activation kinetics and determine the optimal metabolic phenotype and fitness to shape product potency.
Both glycolysis and mitochondrial oxidative phosphorylation shape T cell activation and effector function. Glycolysis drives a robust, short-lived effector phenotype. In contrast, OXPHOS and fatty acid metabolism tunes persistence and sustained T cell memory formation.
Open an early window to detect T cell activation responses within minutes of stimulation by measuring glycolytic extracellular acidification rate (ECAR) in real time. Learn more
NovoCyte and Quanteon flow cytometers
High-performance benchtop flow cytometers designed for all levels of users. For quick and accurate immunophenotyping with up to 25 channels of fluorescence using the most sensitive silicon photomultiplier detector technology on the market. Experience the new standard in flow cytometry.
Monitor T-Cell expansion, compound toxicity, drug effects on tumor cell growth, immune cell behavior under different conditions, and much more.
NovoCyte software has a built-in module for cell proliferation modeling, which is shown in the diagram opposite.?It identifies each generation of cell division, and then calculates the proliferation index.
NovoCyte Quanteon is the most advanced benchtop flow cytometer on the market today. With four lasers and up to 25 independent photomultipliers it is ideal for conducting immuno-oncology research to support the most demanding panels. Add the NovoSampler Q for walk-away automation.
The right shows an 18-color pan-leukocyte immunophenotyping panel on PBMCs using the NovoCyte Quanteon. This panel was designed to monitor different immune subsets with activation and differentiation markers.?
Above, we show a 18-color pan-leukocyte immunophenotyping panel on PBMCs using the NovoCyte Quanteon, adapted from OMIP-024 (Cytometry Part A,
85A:995-998, 2014). This panel was designed to monitor different immune subsets with activation and differentiation markers.
Agilent SureGuide chemically synthesized sgRNAs
Agilent chemically synthesized guide RNAs facilitate the full potential of CRISPR for cellular engineering and immunotherapy.
These long, high quality, chemically synthesized single guide RNAs are ideal for CRISPR-based cell engineering. With customization options, such as modifications to increase stability and specificity, for consistent results and robust, novel therapeutics development. Small scale research grade manufacturing and large-scale GMP manufacturing capabilities result in an end-to-end solution for your cell engineering needs–from research through to clinical trials, providing quality and reliability you can trust.
Proprietary chemistries enable the synthesis of long, high purity sgRNAs. This process facilitates the scalable manufacturing of highly pure sgRNAs, affords unique options for sgRNA design, and precise installation of molecular functionality to augment CRISPR-Cas performance.
Customization options, include the ability to add modifications to increase stability and specificity, for more consistent results. For example, adding a single modification at either position 5 or 11 of the hemoglobin subunit beta (HBB) guide RNA sequence dramatically reduced the level of off-target activity while maintaining a high level of activity at the on-target site.
Accelerate your immune cell therapy development
Want to learn more about the Agilent immune cell therapy tool bench? Schedule a seminar with our product specialists and take the first step in improving cell function and potency.
For Research Use Only. Not for use in diagnostic procedures.