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Unicell based immunocyte typing quantitative analysis method

A technology for quantitative analysis of immune cells, applied in the biological field, to achieve fast analysis speed, avoid mutual influence, and high sensitivity

Pending Publication Date: 2019-11-05
TAIZHOU POLARIS BIOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] The technical problem to be solved by the present invention is to overcome the above-mentioned shortcomings, and the research design is aimed at the characteristics of highly differentiated immune cells, and simultaneously conducts high-throughput analysis and detection of sufficient antigen markers in all types of immune cells at the single-cell level.

Method used

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  • Unicell based immunocyte typing quantitative analysis method
  • Unicell based immunocyte typing quantitative analysis method
  • Unicell based immunocyte typing quantitative analysis method

Examples

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Embodiment 1

[0080] Utilizing the present invention to quantitatively type the immune cell populations of patients with non-small cell lung cancer (NSCLC) in the oncology department of a certain hospital, the method comprises the following steps:

[0081] (1) Peripheral blood samples (7.5 mL) from NSCLC patients were centrifuged and filtered, and then resuspended with 1 mL of phosphate buffer to make a single cell suspension;

[0082] (2) Add 3 mL of erythrocyte lysate to the above single cell suspension, vortex or invert to mix, place on ice for 15 minutes, vortex and mix twice in between, centrifuge at 450g 4°C for 10 minutes to precipitate white blood cells , carefully discard the supernatant, and resuspend the cells in 1 mL of phosphate buffer;

[0083] (3) Add 100 μL of 16% formaldehyde solution to the cell suspension, and fix at room temperature (25° C.) for 10 minutes;

[0084] (4) Wash the cells and resuspend the cells with staining buffer;

[0085] (5) The following metal-labeled ...

Embodiment 2

[0103] Utilize the present invention to quantitatively type the immune cell population of XX Hospital Department of Dermatology systemic lupus erythematosus (SLE) patient, comprising the following steps:

[0104] The analysis sample was 7.5 mL of peripheral blood from SLE patients. The sample processing method and data analysis method were the same as in Example 1. The selected antibody combination was: [1] basic immune cell typing combination + [2] B lymphocyte typing combination.

[0105] Example results see Figure 5 , Figure 6 As shown, each node represents a class of immune cell subsets, the size of the node circle indicates the number of cells, and the gray value of the node circle indicates the normalized median value of CD19 or CD20 protein expression. CD45+, CD19+, CD20+ B cell population.

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Abstract

The invention provides a unicell based immunocyte typing quantitative analysis method. The method comprises that (1) a blood sample or a focus tissue biopsy sample of a patient is collected; (2) an antibody labeled with metal elements is used to mark cells of the sample with multiple targets at the same time; (3) the marked cell sample is detected by a mass spectrum flow type detection system; (4)on the basis of the obtained mass spectrum flow type detection data, different immunocyte subgroups are clustered for analysis, and the expression level of the different subgroup cells in a tumor immune target is quantitatively analyzed; and (5) after data processing and calculation of a dimension reducing algorithm, a 2D point diagram, a SPADE graph or a thermograph are drafted, data is displayed in a visualized way, and an informatics analysis model is constructed to cluster and analyze the different immunocyte subgroups. The rare immunocyte groups and antigen with lower expression are detected sensitively by high-flux analysis detection, the immunization typing analysis is more accurate, and the method has prospects of clinical application.

Description

technical field [0001] The invention relates to biotechnology, in particular to an analysis method of immune cells, in particular to a method for quantitative analysis of immune cell typing based on single cells. Background technique [0002] The human immune system is composed of a large number and very rich types of immune cells. There are two main types of immune cells, lymphocytes and myeloid cells. Among them, bone marrow cells include granulocytes, monocytes, etc., and can be divided into basophils, eosinophils, neutrophils, and monocytes according to the difference in cell shape and function. The stimulators in turn differentiate into various types of macrophages or dendritic cells. Lymphocytes are a type of white blood cells and are the most important cell components involved in the body's immune response. Lymphocytes can be divided into T lymphocytes, B lymphocytes, and natural killer cells according to the source of lymphocyte differentiation, the surface antige...

Claims

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Application Information

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IPC IPC(8): G01N33/68
CPCG01N33/6848G01N33/6893
Inventor 魏然王宇翀孙双午
Owner TAIZHOU POLARIS BIOLOGY CO LTD
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