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Accurate detection kit for typing of tumor immune cell subset

A technology of immune cells and kits, applied in the field of cell biology detection, which can solve the problems of complex experimental operations, high cost, and few types of metal pre-labeled antibodies

Active Publication Date: 2020-12-29
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, the commonly used mass spectrometry flow system includes CyTOF mass spectrometry flow cytometry system produced by Fluidigm company. The detection function of this system is powerful, and there are more than 600 kinds of antibodies that have been metal-labeled for mass spectrometry flow analysis, but there are still metal labels. Disadvantages such as a single source of antibodies, too few types of metal pre-labeled antibodies, high cost, and complicated experimental operations
At present, there is still a lack of detection kits directly used for comprehensive monitoring and rapid and accurate typing of tumor immune cells. Therefore, it is of great significance to develop a detection kit for tumor immune cell subgroup typing based on mass spectrometry flow cytometry technology.

Method used

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  • Accurate detection kit for typing of tumor immune cell subset
  • Accurate detection kit for typing of tumor immune cell subset
  • Accurate detection kit for typing of tumor immune cell subset

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Immune cell typing detection antibody panel

[0063] This example provides a set of antibody panels for the detection of immune cell typing. The antibody panel contains a total of 42 metal pre-labeled antibodies. The antibody panel can identify all T cell subsets (including T, Treg, central memoryT, effector T, effector memory CD4 + T, CD8 + T) as well as B cells, NK cells, granulocytes, monocyte-macrophage subsets, etc., and also covers immune checkpoints and cell proliferation indicators.

[0064] The antibody panel combined with mass spectrometry and flow cytometry technology can be used for the typing and detection of immune cells, and 42 antibodies use 42 channels.

[0065] Table 1 shows the antibody clone numbers, metal element labels and antibody-bound markers of the 42 metal-prelabeled antibodies. Among them, markers numbered 11, 15, 24, and 38 are intracellular proteins, and others are extracellular (cell surface) proteins.

[0066] Table 1 Antibo...

Embodiment 2

[0069] Example 2 Application of Antibody Panel in Immune Cell Typing of Tumor Patients

[0070] In this example, the antibody panel of Example 1 is used to detect the typing of immune cells in tumor patients, and the specific method is as follows:

[0071] 1. Prepare fresh peripheral blood from normal people and tumor patients, and extract PBMC from peripheral blood mononuclear cells.

[0072] 2. Divide the peripheral blood mononuclear cells PBMC extracted in step 1 into two groups, the control group and the experimental group, each group is about 3 × 10 6 Cells were resuspended in PBS to adjust the volume to 1 mL.

[0073] 3. Add cisplatin (Cell-ID Cisplatin, 201094, Fluidigm) at a final concentration of 5 μM, and stain at room temperature for 2 minutes.

[0074] 4. Add 2ml of Cell Staining Buffer (201068, Fluidigm) to terminate the reaction, and centrifuge at 500×g for 5 minutes at room temperature.

[0075] 5. Aspirate the supernatant, add 50 μL blocking solution (5 μl Fc ...

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Abstract

The invention relates to the technical field of cell biology detection, and particularly relates to an accurate detection kit for typing of a tumor immune cell subset. The invention provides markers for single-cell immune cell typing detection and an antibody combination thereof. The combination covers 42 markers and metal pre-labeled antibodies thereof for immune basic detection, lymphoid cell extension detection and myeloid cell extension detection. The antibody combination and a mass spectrum flow cytometry are used for typing detection of immune cells, various detection requirements can bemet, accurate detection of complex tumor immune cell heterogeneity and rapid and accurate typing of the immune cell subset are achieved, single-staining control and calculation compensation of all channels are not needed, samples are effectively saved, and the detection efficiency is improved.

Description

technical field [0001] The invention relates to the technical field of cell biology detection, in particular to a precise detection kit for tumor immune cell subgroup typing. Background technique [0002] The tumor microenvironment contains many immune cells other than tumor cells, and immune cells play a key role in the occurrence and development of tumors and clinical tumor immunotherapy. The tumor microenvironment is affected by immune cells, and the immune status also affects the progression of tumor disease. Whether in the microenvironment of tumor tissue or in the peripheral blood circulation system, immune cells have very important detection significance and clinical diagnostic value. Detecting the status of the immune system of tumor patients can accurately classify tumor patients and guide subsequent treatment. [0003] Most of the analytical methods and technical means at the cellular or molecular level of traditional tumor immunoassays are based on population ce...

Claims

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Application Information

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IPC IPC(8): G01N33/569G01N33/58G01N33/68G01N15/14
CPCG01N33/56966G01N33/58G01N33/6848G01N33/6854G01N15/14G01N15/00G01N15/01
Inventor 纪建国王青松朱文苑费丹霞
Owner PEKING UNIV
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