Chimeric Antigen Receptor Streamlining Solutions
Chimeric Antigen Receptors (CARs) are engineered to specifically target cancer cell antigens and enhance T-cell proliferation and cytotoxicity. Target specificity and T-cell lifecycle are key factors in determining eventual therapeutic success.
After isolation of specific cell types from patient or donor` blood, T-cells are genetically modified ex vivo to express a CAR protein that recognizes specific tumor antigens. The goal of CAR construct engineering and screening is to overcome limitations such us toxicity, low antigen sensitivity and to achieve increased anti-tumor efficacy without compromising safety.
Complex biological interactions between CAR construct and tumor antigen require the utilization of orthogonal in vitro cell and protein analysis assays.
Solutions for CAR Construct Design and Screening
Octet® Label-Free Biomolecular Interaction Analysis Platform
Octet®️ platforms, available using Bio Layer Interferometry (BLI) or Surface Plasmon Resonance (SPR), measures protein-protein interactions in parallel, without the use of detection agents. These robust approaches enable fast, real-time characterization of expressed proteins, even in complex and unpurified samples.
Featured applications include:
- Quantitation of protein expression and antigen density
- Detection and screening of CAR affinity by binding and competition assays
- Epitope binning of antibodies, antibody fragments (ScFVs)
Related White Paper: Streamlining Affinity Analysis for Accelerated Lead Screening
CellCelector Automated Cell Selection and Retrieval Platform
The CellCelector platform is a unique and fully automated cell imaging and retrieval platform developed for detection, selection and isolation of single cells, clusters, spheroids and organoids as well as single cell clones and adherent colonies. In the field of development of CAR-T constructs the platform provides great solutions for:
- Colony picking after genome editing (CRISPR) - Select the cells with the desired genetic characteristics with single cell cloning
- Single Cell Heterogeneity Analysis - Select target cells based on fluorescent markers and through distinct morphological features
- Isolation of a Precise Number of Cells - Isolate the exact number of cells needed for high-quality spike-in reference in validation assays
Related Application Guide: Fully Automated Image-Based Single Cell and Colony Picking for Stem Cells
Incucyte® Live-Cell Analysis Platform
Enables real-time, live-cell imaging within the stable condition of the incubator. Make informed decisions about your cultures, rapidly optimize and improve your workflows, plus study complex live-cell assays to fast track your next discovery!
- Monitoring cell health, proliferation as well as toxicity, viability on model cell system
- Evaluation of protein expression
Related eBook: Harnessing the Power of T Cells
iQue® Advanced Flow Cytometry Platform
Achieve fast, reliable, high-throughput analysis using minimal sample with the iQue® Advanced Flow Cytometry Platform with a portfolio of assays for adherent cells
- Cell health, viability and proliferation of modified T-Cells
- CAR gene, protein expression evaluation
- Antibody | ScFv library screening as most commonly used ligand-binding domains in CAR structures
Related White Paper: Understanding T Cell Phenotype and Function to Enable Improved Therapeutics
Arium® Pro VF or UF Ultrapure Water Systems with Integrated Ultrafilter
T-cell engineering includes different gene editing techniques which are sensitive reactions to nuclease activities. Arium® Ultrapure Water Systems produce ASTM Type 1 water, as well as RNase/DNase and endotoxin-free water to ensure consistent, high-quality results.
- High removal of TOC, Endotoxins, DNAse and RNAse
- Can be operated with various accessories, e.g. printer remote dispenser
- Includes IQ/OQ, preventive maintenance, etc.
- For a daily water consumption of up to 100 L/day.
- Includes visual SOP monitoring with push notifications for alerts and warnings
Related Application Note: Water is Life - Ultrapure Water in Cell Cultivation
Sartorius CAR Construct Design and Screening Solutions Offer:
CellCelector represents the perfect tool for targeted isolation, picking and transfer of genetically modified cells with the desired genotype. The High-Throughput Nanowell-based Image Verified Cloning (HT-NIC) single cell cloning method with integrated monoclonality and viability proof enables industry leading outgrowth rates after clone transfer into 96 or 384 well plates.
Octet® BLI software has a built-in epitope binning module to enable easy assay design, performance, and interpretation allowing for discriminating between antibodies that bind the same antigen but not the same epitope in cross-competition assays.
Relieve capacity bottlenecks throughout your entire experiment with the fastest plate sampling, integrated analysis, and novel data reduction tools. With the iQue® Advanced Flow Cytometry Platform, you can analyze a 96-well plate in as few as 5 minutes or a 384-well plate in as few as 20 minutes for a faster time to result.
Related Publications
- GPC2-CAR T cells tuned for low antigen density mediate potent activity against neuroblastoma without toxicity: Cancer Cell
- The IgG4 hinge with CD28 transmembrane domain improves VHH-based CAR T cells targeting a membrane-distal epitope of GPC1 in pancreatic cancer | Nature Communications
Reference: Rafiq, S., Yeku, O., Jackson, H. et al. Targeted delivery of a PD-1-blocking scFv by CAR-T cells enhances anti-tumor efficacy in vivo. Nat Biotechnol 36, 847–856 (2018). https://doi.org/10.1038/nbt.4195