September 10 – 13, 2019
Seaport Hotel & World Trade Center
Boston, MA USA
Don’t comprise your ambition, visit Sartorius Booth 75 to learn about our innovative solutions to advance CAR-T discovery and development.
We know you need to trust your therapy. We believe your goal is to achieve results that are reproducible, publishable, and actionable, resulting in breakthrough therapy. That’s why we strive to provide innovative solutions for your needs, from biomarker identification to production. We want to ensure you develop the safest CAR-T product, with high levels of productivity, and that you never compromise your ambition.
- Intellicyt iQue®3 - Platform enables high throughput analysis of immune cell profiling with minimal sample volume and rapid results, in real time and without compromising your cell cultures.
- Incucyte® System and Incucyte® Live-Cell Assays - Achieve insights into the cellular processes with reliable, quantitative data. Enjoy real-time continuous automated imaging and analysis of cell health and viability, CAR-T cell homing (chemotaxis), activation (T cell proliferation and clustering), immune cell killing assays, and antibody mediated phagocytosis, in a physiologically relevant environment that does not require you to be in the laboratory for every datapoint.
- Picus® and Tacta® Pipettes - Sample preparation is a delicate process. The integrity of the sample must be maintained so you can trust your results. This is important when working with the patient-derived cultures that will become CAR-T cells and when preparing samples used for in vitro assays to assess immune cell killing, phagocytosis, apoptosis, and more. We offer a variety of solutions to assist with sample preparation.
- The Mycap® CCX Cell Expansion System - supports aseptic cell expansion from vial to bioreactor in shake flasks outside of the biosafety cabinet.
- Microsart® ATMP Rapid Real-time PCR Mycoplasma/Bacterial Detection Kits and Microsart® Mycoplasma qPCR Detection Assays - are a rapid, specific, and sensitive method for contaminant detection during CAR-T cell characterization. Unique and highly specific TaqMan® qPCR assays are validated according to the International Pharmacopeia and increase patient safety prior to treatment. Confidently generate actionable, reproducible results in hours, not days.
At the show, please visit our poster presentation.
Title: Gaining Insight for Personalized T Cell Manufacturing Utilizing High Throughput Flow Cytometry
Authors: Zhaoping Liu, Andrea Gomez-Donart and John O’Rourke
Essen BioScience, Albuquerque, New Mexico, USA.
The bio-manufacturing process of personalized T cell products as live medicine is critical to adoptive T cell therapy. Monitoring the ex vivo activation of human T lymphocytes is key to developing an optimized and scalable T cell manufacturing process. In addition, ex vivo expansion is correlated with the persistence of infused T cells and patient clinical outcomes. Recent clinical studies show a subset of functional memory T cells including T stem cell-like memory cells (Tscm), T central memory cells (Tcm) and other less differentiated T cell subsets are responsible for long-term anti-tumor responses. All these suggests that ex vivo monitoring of T cell activation and T cell expansion are important to significant clinical success in adoptive T cell therapy.
To address the need to rapidly monitor cell phenotype, T cell activation, cell health and function for improved ex vivo activation and expansion protocols in the bio-manufacturing of personalized T cell medicine, we developed a streamlined workflow based on high throughput flow cytometry technology to help generate more critical insight for informative decision-making. As a proof of concept, we built 2 immune panels to functionally profile T cell activation and T memory cells, respectively. Both panels take advantage of multiplex capabilities, assay miniaturization and easy-of-use workflow via high throughput flow cytometry. Each panel uses a cell and bead mixture assay format to simultaneously measure T cell phenotype, cell health, secreted cytokines and functions.
In order to show the workflow of T cell activation monitoring via high throughput flow cytometry, peripheral blood mononuclear cells (PBMCs) from a cohort of 12 donors including healthy, Non-Hodgkin’s lymphoma (NHL), and Chronic Myeloid Leukemia (CML) patients were profiled in a single 96-well plate for T cell activation. Over the course of a three-day activation protocol with CD3/CD28 antibody-coated magnetic beads, 11 secreted cytokines and 13 cellular endpoints were measured within 20 minutes per 96-well assay plate per day on iQue3 platform, a high throughput flow cytometer with 3 lasers, 13 fluorescent channels and an integrated, plate-level based data analysis software named ForeCyt. These results provide donor-specific, time-resolved cellular and secreted cytokine profiles as a T cell activation signature for the decision making in T cell bio-manufacturing. To demonstrate the screening of ex vivo expansion conditions to preferentially grow less differentiated T cells on high throughput flow cytometry technology, PBMCs from 3 healthy donors cultured with 14 different combinations of cytokines including IL-4, IL-6, IL-7, IL-15, IL-21 and IFNβ in a single 96-well plate and were screened on Day 2, 5 and 8 with a multiplexed 10-color T memory panel. We identified T-cell expansion in the media supplemented with the cytokine cocktails containing IFNβ that promoted the frequency of T central memory cells. In addition, the cytokine cocktails containing IL-21 in the culture media increased the frequency of T stem cell-like memory cells. Furthermore, the secretion of IL-10 during T cell expansion correlated with the frequency of T central memory cells. All these data show that high throughput flow cytometry is a robust tool, not only to rapidly monitor day-to-day T cell activation for scalable T cell manufacturing, but also to screen ex vivo culture conditions and media supplementation leading to preferential expansion of desired T cell memory subsets. This study highlights the capability of the iQue3 platform to rapidly characterize multiple cellular and secreted endpoints in a single multiplex assay, and to provide high content data visualization to drive faster decisions in the personalized T cell manufacturing.