Immune Monitoring Services
Comprehensive Characterization of the Immune Response
The Immune Monitoring shared resource provides a full suite of services to monitor and characterize immune responses. We offer support for the development and testing of new CAR T cells, including product-release testing of newly engineered T cells for Phase 1 clinical trials. As part of this effort, our tetramer and molecular assays teams provide services to the Hutch's Therapeutic Products shared resource. The IM team also works with investigators to develop new assays and bring in cutting-edge new technologies.
To request services, please refer to our rates and scheduling page.
Certain instruments are open to all investigators on a first-come, first-served basis and no reservation is currently required.
ON THIS PAGE
Tetramer Production | Cytokine Assays | Molecular Assays | Monitoring and Phenotyping Immune Cells | Molecular Cloning by DNA Assembly | Equipment
Schedule With Us
To schedule services, on-site assistance or training with the Immune Monitoring, or to get more information about how we can work with you, contact the IM team lead:
Jianhong Cao, Ph.D.
Director/Senior Staff Scientist
Immune monitoring specialist Eric Griffiths loads an ELISA assay plate onto the Synergy H4.
Centrifuge in the Immune Monitoring core
Immune monitoring specialist and cytokine lab lead Rick Lawler analyzes data from a Luminex assay.
Immune monitoring specialist Eric Griffiths loads an ELISA assay plate onto the Synergy H4.
Centrifuge in the Immune Monitoring core
Immune monitoring specialist and cytokine lab lead Rick Lawler analyzes data from a Luminex assay.
In a separate production line in a clean room, we produce tetramers in a singular procedure and sterilized them at the end. Access more information and a complete list of the MHC Class-I alleles and fluorochrome-conjugate options available for tetramer production by downloading this PDF:
We use fast protein liquid chromatography, or FPLC, to purify and analyze proteins. Depending on the type of the column used, proteins are separated based on the properties of size, charge and hydrophobicity. Our FPLC instrument is used mainly for pMHC tetramer production. Upon request, we can also run other protein samples. We have three FPLC instruments available, including one reserved only for clinical-grade tetramer production.
This is not generally a shared instrument; please contact our team to inquire about use.
We perform quantitative immunoassays on your biological samples to give you concentrations of various cytokines and other analytes in serum, plasma and cell culture supernatants.
To learn more about our library of common cytokines, as well as other proteins we develop at researchers’ request, please refer to our FAQ:
We offer automated single-cell analysis of cytokines via the IsoLight system from IsoPlexis. The platform is designed to understand and characterize differences among single cells. By mapping thousands of cells per sample, the system can reveal full functional profiles and polyfunctionality among cell subsets to determine patient response and product quality.
Please contact our team to request use of the instrument.
The SR-X Ultra-Sensitive Biomarker Detection System from Quanterix enables ultra-sensitive biomarker detection. The SR-X allows multiplex detection of up to six analytes per well, with low volume requirements to increase productivity and throughput. An extensive menu of Simoa assay kits are available to measure critical biomarkers with substantially higher sensitivity than standard immunoassay methods, enabling detection of both normal and acute levels with high precision across a range of sample types.
Please contact our team to request use of the instrument.
We offer the Agilent 4200 TapeStation for DNA analysis. This includes high-sensitivity analysis of DNA from 100 to 5000 base pairs. Please contact our team to request use of the instrument.
The Qubit 4 Fluorometer from Thermo Fisher Scientific is designed to accurately measure DNA, RNA and protein quantity, as well as RNA integrity and quality, using the highly sensitive Qubit assays.
Please contact our team to request use of the instrument.
Spectratyping is a set of multiplex RT-PCR reactions that amplifies all T-cell receptor beta-chain families to reveal the clonal-length polymorphism in the CDR3 region. This technique is used to access clonal composition in T-cell populations. TCR spectratyping is often used to determine clonal expansion, contraction and deletion during immune response and reconstitution. It can be performed on PBMC samples or FACS-sorted cell subsets.
We aid investigators seeking to monitor and phenotype immune cells according to differentiation, activation, functional markers and other characteristics. Our technologies and capabilities allow users to characterize cell-surface and intracellular proteins, detect antigen-specific T cells, and isolate specific cell subsets for further analysis. We provide flow cytometry panel development, analysis and data presentation in accordance with specific study requirements.
Our capabilities are supported by close connections to other Fred Hutch shared resources, including the Flow Cytometry core. We also work with users to characterize cells stored with the Specimen Processing & Research Cell Bank shared resource.
The robotic BioXp 3200 system workstation builds double-stranded DNA fragments and plasmid clones from custom-designed reagents. Gibson Assembly is the underlying molecular technology. We order designed DNA sequence reagents from CODEX DNA, which will usually be delivered in three to five business days. Our staff will build the DNA overnight on the BioXp.
The following are available:
- dsDNA fragments: DNA fragments between 300 and 1800 base pairs can be assembled. Up to 32 fragments can be assembled in each BioXp overnight run.
- Cloning: DNA fragments can be cloned into your vector of choice. The customer provides the linearized vector (up to four different vectors per run) to our lab and a Gibson Assembly reaction takes place on the BioXp with your assembled fragments. The customer can then transform the potential clone into competent E. coli cells and verify the constructs by sequencing.
- RapidAMP (under development): Eliminate the transformation step by amplifying your clonal plasmid DNA for direct use in transfection. Up to 24 constructs can be built with up to four different vectors in each run.
We offer the Agilent 4200 TapeStation for DNA analysis. This includes high-sensitivity analysis of DNA from 100 to 5000 base pairs. Please contact our team to request use of the instrument.
The Synergy H4 with Hybrid Technology is a multi-mode detector for reading 96-well dishes. The reader has quadruple monochromator-based optics and filters that provide flexibility in usage. Users can select any wavelength, from the low UV to the near-infrared. Detection methods include fluorescence, time-resolved fluorescence, fluorescence polarization, AlphaScreen/AlphaLISA, luminescence and UV-visible absorbance. Read methods include end-point, kinetic, spectral scanning, and well-area scanning. The instrument can read plate formats from six to 384 wells.
This ImmunoSpot reader from CTL is specially designed for reading ELISPOT and FluoroSpot plates. The image-acquiring and analysis system is equipped with a microscope that can scan 96-well plates mounted on a workstation. The accompanying software enables the recognition and quantitation of immunospots in each well. This flexible system can read different plates from various suppliers either automatically or manually, with many options for manipulation.
The DLR-ready Berthold Model Centro LB 960 microplate luminometer has three injectors.
The Roche LightCycler 480 performs quantitative real-time PCR analysis as well as methylation and high-resolution melting analyses.
Hidex Sense is a scintillation and luminescence counter reads 96-well plates. It is commonly used for the 3H-thymidine incorporation lymphoproliferation assay and chromium-51 release assay. It also reads luminescence for many reporter assay systems.
Fast read time, compatibility with both 96- and 384-well plates, and ease of integration with front-end automation systems make the FLEXMAP 3D® the platform of choice for high throughput applications. The platform uses both magnetic and non-magnetic microspheres and is capable of simultaneously measuring up to 500 genes or proteins from a small sample. Automated start-up, shut-down, and maintenance routines make the FLEXMAP 3D easy to use and maintain.
Key Features:
- Multiplexing: Up to 500 analytes in a single sample
- Dynamic Range (Typical): ≥4.5 logs
- Read Time: 96-well plate in ~20 min (up to 144,000 tests/hour); 384-well plate in ~75 min (up to 153,600 tests/hour)
- Daily Start-Up/Shut-Down: ~30 min
This is not generally a shared instrument; please contact our team to inquire about use.
The z-Movi® is a unique instrument that measures the avidity between immune cells and their targets, enabling you to identify the most potent immunotherapeutic effector cells. This new technology provides you with predictive, reproducible, and fast high-throughput results at a single-cell resolution without compromising cell viability. All within a compact little box that easily fits inside the flow hood for sterile and safe sample handling.
The SR-X Ultra-Sensitive Biomarker Detection System from Quanterix enables ultra-sensitive biomarker detection. The SR-X allows multiplex detection of up to six analytes per well, with low volume requirements to increase productivity and throughput. An extensive menu of Simoa assay kits are available to measure critical biomarkers with substantially higher sensitivity than standard immunoassay methods, enabling detection of both normal and acute levels with high precision across a range of sample types.
This is not generally a shared instrument; please contact our team to inquire about use.
The Qubit 4 Fluorometer from Thermo Fisher Scientific is designed to accurately measure DNA, RNA and protein quantity, as well as RNA integrity and quality, using the highly sensitive Qubit assays.
Please contact our team to request use of the instrument.
The robotic BioXp 3200 system workstation builds double-stranded DNA fragments and plasmid clones from custom-designed reagents. Gibson Assembly is the underlying molecular technology. We order designed DNA sequence reagents from CODEX DNA, which will usually be delivered in three to five business days. Our staff will build the DNA overnight on the BioXp.
The following are available:
- dsDNA fragments: DNA fragments between 300 and 1800 base pairs can be assembled. Up to 32 fragments can be assembled in each BioXp overnight run.
- Cloning: DNA fragments can be cloned into your vector of choice. The customer provides the linearized vector (up to four different vectors per run) to our lab and a Gibson Assembly reaction takes place on the BioXp with your assembled fragments. The customer can then transform the potential clone into competent E. coli cells and verify the constructs by sequencing.
- RapidAMP (under development): Eliminate the transformation step by amplifying your clonal plasmid DNA for direct use in transfection. Up to 24 constructs can be built with up to four different vectors in each run.
We offer automated single-cell analysis of cytokines via the IsoLight system from IsoPlexis. The platform is designed to understand and characterize differences among single cells. By mapping thousands of cells per sample, the system can reveal full functional profiles and polyfunctionality among cell subsets to determine patient response and product quality.
Please contact our team to request use of the instrument.
We use fast protein liquid chromatography, or FPLC, to purify and analyze proteins. Depending on the type of the column used, proteins are separated based on the properties of size, charge and hydrophobicity. Our FPLC instrument is used mainly for pMHC tetramer production. Upon request, we can also run other protein samples. We have three FPLC instruments available, including one reserved only for clinical-grade tetramer production.
This is not generally a shared instrument; please contact our team to inquire about use.
The Luminex® 100/200™ System is a flexible analyzer based on the principles of flow cytometry that enables you to multiplex (simultaneously measure) up to 100 analytes in a single microplate well, using very small samples. The system delivers fast and cost-effective bioassay results on many assay formats including nucleic acid assays, receptor-ligand assays, immunoassays, and enzymatic assays.
Please contact our team to request use of the instrument.