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 T cells therapy for 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.
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.
Senior Director/Senior Staff Scientist
ON THIS PAGE
Tetramer Production | Cytokine Assays | Molecular Assays | Monitoring and Phenotyping Immune Cells | Equipment
Immune Monitoring Specialist Duy working on cell analysis by flow cytometry.
FLEXMAP 3D in the Immune Monitoring core
Immune monitoring specialist and cytokine lab lead Rick Lawler performing cytokine analysis.
Immune Monitoring Specialist Duy working on cell analysis by flow cytometry.
FLEXMAP 3D in the Immune Monitoring core
Immune monitoring specialist and cytokine lab lead Rick Lawler performing cytokine analysis.
We produce Class-I tetramers and monomers for research use. Recombinant Class-I heavy chain and β2-microglobulin are expressed in E. coli. The MHC monomeric complex is refolded with epitopic peptides in vitro. After biotinylation, monomers are conjugated to streptavidin to form tetramers.
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 are now offering limited MHC Class-II tetramer production. Please contact lab staff for more information.
We produce tetramers with a DNA barcode, which can be used for various single-cell sequencing applications, including CITE-seq. Please contact our team to request use of the instrument.
Certain Class-I tetramers will be produced without peptide. Alternatively, we can also produce certain tetramers with UV-cleavable peptides.
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.
The unique feature of each T-cell clone is found in its recombined T-cell receptor sequences. We can identify the clonal TCR a-chain sequence by real-time PCR and direct sequencing of the clonal cell sample. The clonal TCR-a/β sequence can be cloned and used for quantitative PCR analysis.
With knowledge of the clonal TCR sequence, specific primers and a probe for quantitative RT-PCR analysis can be designed, allowing the quantitation of the clonal T cells in a cell population. We provide this service in the setting of adoptive T-cell transfers in immunotherapy trials for cancer patients. We also provide custom-designed qPCR assays for CAR T-cell therapy trials.
The immunoSEQ Assay from Adaptive Biotechnologies uses a multiplex PCR-based assay to sequence directly from genomic DNA. immunoSEQ provides accurate, quantitative and high-throughput sequencing of the CDR3 variable chain of immune-cell receptors.
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.
The SMARTer Human TCR a/b Profiling enables users to analyze T-cells receptor diversity from human RNA samples or directly from cells. The kit generate data for both alpha and beta chain diversity. Libraries produced with the kit are indexed and ready for sequencing on Illumina platform.
The kit uses switching mechanism at 5’ end of RNA templates, full-length cDNA synthesis technology, and pairs NGS with a 5’-RACE approach to capture the complete V(D)J variable regions of T cell receptor Alpha and Beta genes.
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.
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 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 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.
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 Roche LightCycler 480 performs quantitative real-time PCR analysis as well as methylation and high-resolution melting analyses.
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.