||Main Cell Type
||Identifies all T-cells and is the most specific marker for T-cells, including lineage based based markers such as CD4 and CD8.
||Indicates that the T-cell has previously encountered its cognate antigen. An increase in CD45RO+ T-cells in tumors is associated with a better prognosis.
||A marker of T-cell exhaustion indicating that normal function of the cell has been disrupted. May also indiate an immuno-suppressive phenotype. Increased levels of PD-1+ T-cells is associated with a poorer prognosis. PD-1 is the target of multiple therapeutics.
||The kit may be used to interrogate many tumor sample types for Research. A cocktail of pan-cytokeratin and Sox10 is provided. Cytokeratins are expressed in cells of an epithelial origin including most carcinomas. Sox10 is expressed in cells derived from the neural crest including melanocytes that give rise to melanomas.
- The antibody panel content of Ultivue’s kits is defined from voice-of-customer inputs, from collaborators, prospective customers and key opinion leaders.
- Ultivue's product development process begins with the screening and careful selection of antibody clones for each target. Clones are selected for their target specificity, reliability, and widespread acceptance in clinical labs.
- Once selected antibodies have been conjugated to unique DNA barcodes, they are tested to verify that the antibody conjugates retain the same performance as the unconjugated material. This is carried out by comparing the staining quality with gold standard methods and certifying the staining by a qualified reviewer.
- To establish multiplexed panels, a titration series for each antibody conjugate is carried out in FFPE tissue control samples to select optimal antibody staining concentrations. Assay protocol steps are optimized as needed.
- When applicable, the analytical sensitivity of the assay for a given target is tested using standards with a known concentration gradient, such as cell pellet controls.
- To confirm reproducibility specifications, functional testing of the entire kit is accomplished via inter-instrument, inter-day, inter-analyst, and intra-run testing for all protocols. Samples are imaged on a wide range of microscopes and slide scanners from leading suppliers.
- Images from reference samples are formally reviewed by consulting pathologists. Alpha and/or Beta pre-production kits are evaluated by prospective customer labs to ensure the assay meets performance expectations and that the kit documents and instructions are clear and complete.
- Multiple stability tests are run for every component of the kit including real-time, accelerated, in-use, on-board, shipping, and freeze-thaw cycle stability.
- The kits for immuno-profiling are designed to offer seamless integration into standard immunohistochemistry workflows. The optimized manual and automated protocols enable a rapid path from sample to answer in a single day.
- The kit is configured to offer an easy-to-use, fast, and fully automated tissue staining solution using the Leica BOND RX® system. The reagents enable the detection of up to four markers and a nuclear counterstain on whole slides using fluorescence tissue scanners such as the Leica Aperio VERSA® amongst other validated imaging solutions. The resulting multiplexed images are compatible with most digital pathology image analysis solutions, such as the IndicaLabs Halo® platform, enabling the automated quantification of spatial and cellular expression data over whole tissue sections.
- As an example, our kits enable the automated staining of four targets and nuclear counterstain on up to 30 slides at one time on the Leica BOND RX®, in 5 hours. Using current imaging systems, a 2 cm2 FFPE section can be imaged in five fluorescent channels in less than 15 minutes, enabling a true sample to answer workflow within a work day.
A deidentified retrospective sample from a 69 year old, retired female presented with a unilateral breast skin lesion. According to the demographic and outcome information for the sample, the woman was slightly obese and suffered from essential hypertension marked by ischemic heart disease and heart failure. She reported to have never smoked and to have never consumed alcoholic beverages. Menopausal status and previous pregnancies were unknown. Upon excision of the skin lesion, a pathological diagnosis of nodular melanoma was rendered with a UICC stage of T4aNXM0, a Clark level of IV, and a Breslow thickness of 14 mm. The tumor size was 1.5 cm. No lymph nodes were excised or examined.
|PD-1 case study
- Tumor appeared to be inflamed as supported by the following observations:
- Much greater numbers of CD45RO+ cells than PD-1+ cells
- Most of PD-1+ cells were contained in lymphoid-like structures
- Infiltrating T-cells appeared to be memory T-cells
- Further sample analysis could possibly characterize an immunosuppressive or an immunoreactive phenotype