Immuno-oncology Product Resource Guide
Your comprehensive guide to tools for immuno-oncology research.
Our Goal
To support your immuno-oncology research with a comprehensive range of tools and technologies that maximize your time, budget, and data—helping to accelerate your path to discovery and translation to a clinical application.
This guidebook provides educational resources and solutions for immuno-oncology research approaches, including checkpoint inhibition, CAR T cell therapy, and cancer vaccine research. Learn about capabilities, from leveraging innovative products and techniques to time-saving workflow applications.
Contents
- Immuno-oncology review
- Key questions about I-O
- General I-O workflow
- Solutions by approach
- Solutions for checkpoint inhibition
- Solutions for CAR T cell therapy
- Solutions for cancer vaccine research
- Characterize and verify
- Targeted genetic analysis
- Targeted gene expression analysis
- Cell analysis
- Protein analysis
- Ordering information
Immuno-oncology Review
What is I-O?
Immuno-oncology (I-O), also known as cancer immunotherapy, is a rapidly growing field that studies the ways in which the body's own immune system can fight cancer.
Why does I-O research matter?
I-O research aims to develop cancer immunotherapies that go beyond traditional methods such as surgery, chemotherapy, and radiation, by enabling the adaptive immune system to recognize and specifically attack cancer cells while leaving healthy ones undamaged. I-O research can potentially uncover ways to enable immunogenicity of all types of cancer and facilitate long-lasting, protective immunity against future recurrence. Recent breakthroughs in checkpoint inhibition, chimeric antigen receptor (CAR) T cell therapy, and cancer vaccines illuminate the full capabilities of the immune system and how it may be harnessed to combat cancer.
While every immune cell has a key part to play in the landscape of I-O, T cells and T cell-mediated responses are the focal points of I-O research today.
What are some of the promising areas in I-O research?
This handbook provides an overview of three currently trending I-O research areas: checkpoint inhibition, CAR T cell therapy, and cancer vaccines. Figure 1 illustrates their correlation within the cancer-immunity cycle.
Figure 1: Stages of the adapted cancer-immunity cycle can be impacted by I-O approaches such as checkpoint inhibition, CAR T cell therapy, and cancer vaccines. The diagram shows T cells infiltrating tumor tissue, interacting with tumor cells, and the roles of APCs and cancer vaccines in initiating and modulating the immune response.
Checkpoint inhibition
Immune checkpoints are cell pathways crucial in maintaining a normal immune response and protecting tissues from damage when the immune system is activated. Cancer cells dysregulate immune checkpoints and use them as a mechanism of immune resistance. Understanding the interactions between tumor and immune cells is key to I-O research. Natural mechanisms regulate T cell activity via T cell receptor (TCR) interactions. For example, PD-1/PD-L1 is a coinhibitory pathway that 'masks' cancer cells from T cell recognition, preventing T cell attack. Antibodies targeting the PD-1/PD-L1 pathway suppress this function, allowing T cells to recognize cancer cells and initiate cytotoxic activity.
The B7/CTLA-4 pathway plays a role during T cell priming by an antigen-presenting cell (APC). Blocking the CTLA-4 receptor with an antibody allows T cell activation, leading to an anticancer immune response. Multiple costimulatory and coinhibitory receptor-ligand interactions between APCs and T cells are involved in T cell activation or suppression.
Figure 2: Illustrates multiple costimulatory and coinhibitory receptor-ligand interactions between APCs and T cells. It highlights the B7-CD28 family and TNF family members involved in regulating T cell activation.
Adoptive Cell Therapy (ACT) and CAR T Cell Therapy
ACT targets the immune system to fight cancer, rather than targeting cancer directly. This is achieved by genetically modifying a subject's own T cells to target antigens selectively expressed on cancer cells. Successful applications include Tumor-infiltrating lymphocytes (TILs) and CAR T cells, which are engineered to express chimeric antigen receptors for targeting specific cancer cells.
Figure 3: Depicts CAR T cell engineering, involving modification of an individual's T cells to target specific cancer cell antigens.
Cancer Vaccines
Cancer vaccines are another important I-O research area aimed at enabling the immune system to recognize cancer as a threat. This antigen-based method relies on the immune system's ability to recognize proteins to induce an immune response. Researchers identify tumor-associated antigens, called neoantigens, released within the tumor microenvironment to understand tumor formation and spread, informing vaccine development. Dendritic cell (DC) therapy involves activating extracted DCs with tumor fragments, turning them into APCs to induce a secondary immune response. Combination therapies and personalization methods are also being studied.
General I-O Workflow
Thermo Fisher Scientific offers research platforms and products to understand the interplay between the immune system and cancer, accelerating the development of cancer immunotherapies. The workflow starts with biomarker discovery, continues through research on targets, contextual studies within model systems, and concludes with characterization and verification. This workflow applies to checkpoint inhibition, CAR T cell therapy, and cancer vaccine research.
Figure 4: A workflow diagram showing the phases of I-O research: Biomarker discovery, Targets, Modification of model systems, Characterization and verification, leading to Clinical immunotherapy.
Key Approaches:
- Checkpoint Inhibition: Find workflow tools for genomic biomarker discovery, verification, and protein expression, as well as immune checkpoint analysis.
- CAR T Cell Therapy: Find tools spanning the workflow, from T cell isolation and activation to cell engineering and expansion.
- Cancer Vaccines: Find transfection solutions, gene-to-protein services, and genomic biomarker tools.
Solutions for Checkpoint Inhibition
Identifying and validating predictive biomarkers for checkpoint immunotherapy is crucial for optimizing therapeutic benefit, minimizing toxicity, and guiding combination therapy. Solutions include genomic biomarker discovery, verification, protein biology, and cell analysis products.
Figure 1 (re-contextualized): Illustrates the cancer-immunity cycle, highlighting the role of checkpoint inhibition.
Genomic Biomarker Discovery and Verification Tools
- Applied Biosystems™ Clariom™ D Pico assays: Provide transcriptome analysis for novel biomarker discovery, analyzing RNA and alternative splicing events.
- Applied Biosystems™ Clariom™ S Pico assays: Offer gene-level analysis of well-annotated genes for pathway identification and sample screening.
- Applied Biosystems™ TaqMan™ Array Human Immune Response Plate: Enables quantitative gene expression analysis of immune system functions.
- Applied Biosystems™ TaqMan™ Array Human Immune Panel: Provides quantitative gene expression analysis of key immune response gene targets.
- Applied Biosystems™ TaqMan™ Gene Expression Assays: Gold standard for real-time PCR gene expression analysis for biomarker verification.
- Ion Torrent™ Oncomine™ Immune Response Research Assay: A targeted NGS assay for measuring genes involved in tumor-immune interactions and identifying biomarkers.
Find out more at thermofisher.com/checkpointinhibitor
Protein Expression
Straight from the scientist: A quote highlights the ease of use and analytical capabilities of transcriptomics analysis using Clariom assays.
Did you know?: Gibco™ ExpiSf™ Expression System is mentioned as a high-yield, chemically defined baculovirus expression system.
- Gibco™ Expi293™ Expression System: A rapid, high-yield system for protein production from recombinant 293 cells, delivering up to 6x more protein.
- Gibco™ ExpiCHO™ Expression System: Offers higher protein yields and cost savings compared to other transient systems, with scalable protocols.
Figure 5: Demonstrates the scalability of the ExpiCHO system, showing direct scalability from 125 mL to 2 L flask sizes.
Antibodies
Invitrogen™ antibodies: Detect key checkpoint targets with validated primary, secondary, and custom antibodies.
- Includes antibodies for FOXP3, TIGIT, LAG3, ARG1, IDO, NOS2, Granzyme B, and others.
Invitrogen™ Flow Cytometry Panel Builder: A tool for designing multicolor flow cytometry panels with multiple I-O antibodies.
Did you know?: Invitrogen™ antibodies undergo a rigorous two-part testing approach.
Figure 6: Shows immunocytochemical fluorescence analysis of DR3 expression in Ramos cells, with different channels for DR3, nuclei, and F-actin.
Figure 7: Displays immunocytochemical fluorescence analysis of IDO expression in HeLa cells, showing IDO1 protein localization.
Other Assay Tools
- Invitrogen™ CellTrace™ assays: For proliferation measurements based on DNA synthesis or cellular metabolism.
- Invitrogen™ Click-iT™ EdU assays: For analyzing DNA replication in proliferating cells.
- Invitrogen™ PrimeFlow™ RNA assays: For RNA and protein expression analysis.
Figure 8: Presents a Western blot of IDO expression in different cell lines.
Quantitative Protein Analysis
ELISAs, Invitrogen™ ProQuantum™ immunoassays, and Luminex® multiplex platforms are used for quantitative assessment of soluble proteins.
- Invitrogen™ ProcartaPlex™ Immuno-Oncology Checkpoint Markers Panel: For multiplexed checkpoint analysis.
- Invitrogen™ QuantiGene™ Plex Assay: Utilizes Luminex xMAP bead technology for multiplexing and accurate RNA measurement.
Straight from the scientist: A quote from Lisa Butterfield, PhD, highlights the benefits of ProcartaPlex immunoassays for multiplex protein quantitation.
Solutions for CAR T Cell Therapy
Analyzing the immune repertoire to capture TCR diversity aids progress in I-O research. Solutions span workflow from engineering to sequencing.
Figure 1 (re-contextualized): Illustrates the cancer-immunity cycle, highlighting CAR T cell therapy.
Sequencing
- Next-generation sequencing (NGS) using the Ion GeneStudio™ S5 System series: Enables targeted NGS applications with speed and scalability.
- Ion Torrent™ Oncomine™ TCR Beta-LR Assay: A long-read NGS assay for TCR beta chain analysis.
- Sanger sequencing with the Applied Biosystems™ SeqStudio™ Genetic Analyzer: For TCR sequencing and repertoire diversity assessment.
Engineering
Gibco™ LV-MAX™ Lentiviral Production System: A cost-effective and scalable platform for lentiviral vector production, offering higher viral titers and cost reduction compared to PEI-based methods.
Figure 9: Compares lentiviral production methods and shows cost savings with the LV-MAX system.
- Invitrogen™ TrueGuide™ Synthetic gRNA: Ready-to-transfect sgRNA for genome editing.
- Invitrogen™ TrueCut™ Cas9 proteins: For high editing efficiency in gene targets.
- Invitrogen™ Neon™ Transfection System: A device for efficient transfection of primary cells and difficult-to-transfect cells.
- Invitrogen™ GeneArt™ Genomic Cleavage Detection Kit: A PCR-based method to measure genome editing efficiency.
- Invitrogen™ GeneArt™ Gene-to-Protein Services: For fast, reliable protein expression and production.
- Gibco™ Human Plasma-Like Medium: For growth and expansion of T cells.
Figure 10: Demonstrates high-efficiency functional knockout in human primary T cells using TrueCut Cas9 Protein v2 and TrueGuide Synthetic gRNA.
Figure 11: Shows fewer off-target events in human primary T cells with TrueCut HiFi Cas9 Protein.
Translate Your Cell Therapy to the Clinic with CTS Products
Gibco™ Cell Therapy Systems (CTS™) products are cGMP-manufactured media, reagents, and instruments designed for cell therapy applications, facilitating a seamless transition from research to commercialization.
- Isolation and Activation:
- Gibco™ CTS™ Rotea™ Counterflow Centrifugation System: A versatile, closed cell processing system.
- Gibco™ CTS™ Dynabeads™ CD3/CD28: Magnetic beads for T cell isolation, activation, and expansion.
- Engineering:
- Gibco™ CTS™ LV-MAX™ Lentiviral Production System: For cost-effective and scalable lentiviral vector production.
- Gibco™ CTS™ Xenon™ Electroporation System: A closed, scalable platform for electroporation.
- Gibco™ CTS™ TrueCut™ Cas9 Protein: GMP-manufactured Cas9 protein for high editing efficiency.
- Expansion:
- Gibco™ CTS™ OpTmizer™ T Cell Expansion Serum-Free Medium (SFM): For growth and expansion of human T lymphocytes.
- Gibco™ CTS™ AIM V™ Medium: A defined, serum-free formulation for cell proliferation.
- Gibco™ CTS™ GlutaMAX™ Supplement: For improved growth efficiency.
- Gibco™ CTS™ Immune Cell Serum Replacement: For expansion of in vitro-cultured human T cells.
- Wash and Cryopreservation:
- Gibco™ CTS™ Rotea™ Counterflow Centrifugation System: For cell processing applications.
- Gibco™ CTS™ DPBS: A chemically defined medium for cell integrity.
Figure 12: Illustrates gene editing efficiency using Neon and CTS Xenon transfection systems.
Solutions for Cancer Vaccine Research
Cancer vaccine research is evolving, showing promise beyond conventional treatments. Solutions are available to facilitate this field.
Figure 1 (re-contextualized): Illustrates the cancer-immunity cycle, highlighting cancer vaccines.
Genomic Biomarker Discovery and Verification Tools
- Applied Biosystems™ OncoScan™ CNV Assay: Microarray-based assay for whole-genome copy number analysis in solid tumors.
- Applied Biosystems™ CytoScan™ HD Suite: Microarray-based assay for hematological malignancy samples.
Gene Synthesis Services
- Invitrogen™ GeneArt™ Gene Synthesis: For sequence optimization, gene synthesis, and protein expression services.
Engineering
- Invitrogen™ PureLink™ Expi Endotoxin-Free Maxi Plasmid Purification Kit: For endotoxin-free plasmid DNA isolation.
- Invitrogen™ Lipofectamine™ 3000 Transfection Reagent: For high transfection efficiency.
- Invitrogen™ MEGAscript™ T7 Transcription Kit: For high RNA yield.
- Invitrogen™ TransfectionSelect™ product selection tool: Helps select transfection solutions.
Figure 13: Shows how GeneArt expression optimization combined with Gibco systems leads to higher protein expression yields.
White paper: Offers insights on optimizing protein expression for antibody production.
Characterize and Verify
Enhance analysis of genes, proteins, and cells to understand mechanisms of action.
Targeted Genetic Analysis
Confirm and quantify genetic changes for selected DNA targets.
Isolate high-quality genomic DNA or viral DNA: Various kits and reagents are available, including Invitrogen™ DNAzol™, PureLink™, Thermo Scientific™ KingFisher™, and Applied Biosystems™ MagMAX™ kits.
Methods include: Endpoint PCR, Real-time PCR, Sanger sequencing, and Next-Generation Sequencing (NGS).
Key product families: Applied Biosystems™ SimpliAmp™, VeritiPro™, ProFlex™ thermal cyclers, TaqMan™ assays, Ion Torrent™ Oncomine™ assays, Ion GeneStudio™ S5 systems.
Targeted Gene Expression Analysis
Confirm and quantify RNA targets at the gene, exon, or noncoding RNA level.
Isolate high-quality cellular RNA, viral RNA, or miRNA: Solutions include Invitrogen™ RNAlater™, RNaseZap™, TRIzol™, PureLink™ RNA purification kits, and mirVana™ miRNA isolation kits.
Methods include: RT-PCR, Real-time RT-PCR, and NGS.
Key product families: Applied Biosystems™ TaqMan™ Gene Expression Assays, Ion AmpliSeq™ Panels, Ion Torrent™ Oncomine™ assays.
Solution Spotlight: Genetic Analysis Tools
- Ion Torrent™ Oncomine™ Immune Response Research Assay: Monitors the tumor microenvironment (TME) for biomarkers and mechanisms of action.
- Ion Torrent™ Oncomine™ Tumor Mutation Load (TML) Assay: Analyzes cancer-driven genes for mutation counts.
- Ion Torrent™ Oncomine™ TCR Beta-LR Assay: Captures TCR beta chain regions for biomarker discovery and T cell characterization.
- Ion Torrent™ Oncomine™ TCR Beta-SR Assay: Interrogates the CDR3 region of the TCR beta chain for immune status and MRD detection.
- Ion GeneStudio S5 systems: Fast and easy NGS for DNA to data analysis.
- Applied Biosystems™ TaqMan™ Gene Expression Assays: Widely used for gene expression quantification.
Cell Analysis
Understand the impact of perturbations on cellular phenotypes and behavior.
- Observe: Transmitted-light systems for cell, colony, and tissue identification (e.g., Invitrogen™ EVOS™ Imaging Systems).
- Count: Automated counting and viability studies (e.g., Invitrogen™ Countess™ Automated Cell Counter).
- Visualize: Multicolor fluorescence detection and imaging (e.g., Invitrogen™ EVOS™ FL Imaging System).
- Analyze: Flow Cytometry: High-resolution multicolor analysis (e.g., Invitrogen™ Attune™ CytPix™ Flow Cytometer).
- Analyze: Quantitative Imaging: High-content screening and analysis (e.g., Thermo Scientific™ CellInsight™ Systems).
Figure 14: Shows photographic evidence of engineered cell potency and immune synapse formation between CAR T cells and Ramos cells.
Solution Spotlight: EVOS M7000 Imaging System
Provides multicolor cellular imaging, offering insights into spatial relationships and cellular readouts like hypoxia. The system allows precise control of oxygen levels for evaluating cellular responses to hypoxia.
Solution Spotlight: Bigfoot Spectral Cell Sorter
Offers high throughput, safety, and spectral sorting capabilities for applications in immunology, cell and gene therapy research, gene editing, and gene sequencing.
Solution Spotlight: Attune CytPix Flow Cytometer
Provides high-performance capabilities for I-O research, including a high-speed camera for event imaging, structural feature highlighting, and simplified sample preparation. CytKick Autosamplers enable walkaway automation.
Solution Spotlight: CellInsight CX7 LZR Pro High Content Screening (HCS) Platform
Utilizes cell-based imaging screens to identify mechanisms of action in I-O research, assessing cell health and cytotoxicity.
Figure 15: Demonstrates cell painting multiparameter phenotyping using the platform.
Protein Analysis
Analyze the identity, function, and level of expression of key proteins.
- Isolation and Cleanup: Includes chromatography media, dialysis products, desalting products, concentrator devices, and magnetic beads.
- Separate: Features precast protein gels, pour-your-own gel systems, protein ladders, gel tanks, and power supplies.
- Transfer: Offers buffers, membranes, and transfer systems.
- Detect: Includes Western blotting reagents, chemiluminescent substrates, and imaging systems.
- Quantify: Provides total protein and immunoassay solutions, and mass spectrometry reagents.
- Modify: Offers protein crosslinkers and bioconjugation reagents.
Realize the full potential of your protein studies: Enhance western blotting throughput, reproducibility, and sample information.
Ordering Information
A range of products are available for immuno-oncology research, including assays, reagents, instruments, and media. Key categories include:
- Genomic Biomarker Assays: Clariom D/S Pico assays, CytoScan HD Array, OncoScan CNV Assay, TaqMan Arrays.
- Gene Editing & Synthesis: GeneArt Genomic Cleavage Detection Kit, TrueCut Cas9 Protein, TrueGuide Synthetic gRNA, GeneArt Gene Synthesis Services.
- Cell Therapy Systems (CTS): LV-MAX Lentiviral Production System, CTS Immune Cell Serum Replacement, CTS OpTmizer T Cell Expansion Medium, CTS Dynabeads, Xenon Electroporation System.
- Flow Cytometry: Attune NxT/CytPix Flow Cytometers, Bigfoot Spectral Cell Sorter, Click-iT EdU assays.
- Imaging Systems: EVOS Imaging Systems (M7000, M5000, FLoid, XL Core).
- Protein Analysis Tools: ProQuantum immunoassays, QuantiGene Plex Assay, Pierce reagents, Gibco protein expression systems (Expi293, ExpiCHO).
- Sequencing: Ion GeneStudio S5 Systems, Oncomine assays (Immune Response, TML, TCR Beta-LR/SR).
Specific catalog numbers and quantities are available for each product. Further details can be found at the respective product pages linked within the document.
References
The document includes a list of references supporting the information presented.
