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Antibody composition and its application in screening myeloid diseases and detecting immune checkpoints

An antibody composition and immune checkpoint technology, applied in biological testing, disease diagnosis, measuring devices, etc., to achieve the effect of improving the overall survival rate

Active Publication Date: 2021-06-22
信纳克(北京)生化标志物检测医学研究有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] After investigation, there is no report on the FCM technology for the simultaneous detection of myeloid tumors, tumor stem cells, vulnerable cell populations, immune function, and immune checkpoints.

Method used

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  • Antibody composition and its application in screening myeloid diseases and detecting immune checkpoints
  • Antibody composition and its application in screening myeloid diseases and detecting immune checkpoints
  • Antibody composition and its application in screening myeloid diseases and detecting immune checkpoints

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0109] The preparation of embodiment 1 reagent

[0110] The antibody combinations used in this example are:

[0111] Anti-CD15-FITC antibody, anti-CD96-PE antibody, anti-CD33-PE-Dazzle594 antibody, anti-CD34-PE-Cyanine7 antibody, anti-CD117-PE-Cy5 antibody, anti-CD9-PerCP-Cy5.5 antibody, anti-CD45- PerCP antibody, anti-CD38-PerCP-eFluor710 antibody, anti-HLA-DR-APC antibody, anti-CD13 APC-Cy7 antibody, anti-CD19-BV421 antibody, anti-CD4-AF532 antibody, anti-CD36-BV605 antibody, anti-CD7-BV480 antibody, Anti-CD371-BB515 antibody, anti-CD11c-AF700 antibody, anti-CD11b-BV570 antibody, anti-CD200-AF647 antibody, anti-CD14-Pacific Blue antibody, anti-CD56-BV750 antibody, anti-CD71-BV650 antibody, anti-CD2-BV510 antibody, anti- For CD123-BV785 antibody and anti-CD64-BV711 antibody, the above 24 kinds of monoclonal antibody reagents were mixed in the first container according to the volume ratio described in Table 1.

[0112]The antibodies in this example are commercially available...

Embodiment 2

[0114] Example 2 Processing of Specimen

[0115] According to the result of cell counting, add the bone marrow sample anticoagulated with heparin or EDTA into flow tube 1, and ensure that the amount of cells added is about 2×10 6 Add 24 kinds of membrane monoclonal antibody reagents labeled with different fluorescein according to Table 1, mix well with the cell suspension, incubate at room temperature and avoid light for 15 minutes, add 3ml 1× hemolysin, and incubate in the dark Lyse the red blood cells for 10 minutes, centrifuge at 1500rpm for 5 minutes to remove the supernatant, add 3ml PBS buffer to wash, remove the supernatant after centrifugation, and resuspend the cells with 0.5ml PBS buffer, which is the processed specimen and can be tested on the machine.

[0116] Table 1 Antibody mixture and dosage

[0117]

Embodiment 3

[0118] Example 3 Detection and Analysis of Samples

[0119] The specimens processed in Example 2 were detected on a 38-channel flow cytometer with 3 lasers from Cytek Corporation of the United States. After obtaining 100,000 cells per tube, the data was analyzed using a variety of flow cytometry software: spectroflow, flowjo, and kaluza.

[0120] The process of flow cytometry detection and analysis on the machine includes the use of two-dimensional point diagrams and dimensionality reduction diagrams.

[0121] Two-dimensional point diagram analysis process and detection and analysis indicators:

[0122] (1) Use the forward scatter area (forward scatter area, FSC-A) and forward scatter height (forward scatter height, FSC-H) to set the P1 gate to remove the adherent cells, and the P1 gate is a single cell .

[0123] (2) For the cells in the P1 gate, use the FSC-A and the side scatter area (SSC-A) to set the living cell gate (P2 gate).

[0124] (3) Analyze the cells in the P2 ...

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Abstract

The invention provides an antibody composition and its application in screening myeloid diseases and detecting immune checkpoints. The antibody composition comprises anti-CD15 antibody, anti-CD96 antibody, anti-CD33 antibody, anti-CD34 antibody, anti-CD117 antibody, anti-CD9 antibody, anti-CD45 antibody, anti-CD38 antibody, anti-HLA-DR antibody, anti-CD13 antibody, anti-CD19 Antibody, anti-CD4 antibody, anti-CD36 antibody, anti-CD7 antibody, anti-CD371 antibody, anti-CD11c antibody, anti-CD11b antibody, anti-CD200 antibody, anti-CD14 antibody, anti-CD56 antibody, anti-CD71 antibody, anti-CD2 antibody, anti-CD123 antibody and Anti-CD64 antibody. The antibody composition of the present invention can be applied to screening myeloid diseases and detecting immune checkpoints.

Description

technical field [0001] The invention relates to an antibody composition and its application in screening myeloid diseases and detecting immune checkpoints, specifically including a composition containing 24 kinds of antibodies and its application in revealing normal myeloid development, diagnosis of myeloid tumors, and research on tumor stem cells , Targeted therapy target screening, vulnerable cell group research, immune function and application in immune checkpoint detection, belonging to the technical field of blood disease detection. Background technique [0002] Myeloid neoplasms are a group of complex diseases, which can be divided into myelodysplastic syndromes (MDS) according to the proportion of primitive cells, the sublineages involved by malignant clones, the predominant proliferation or pathological hematopoiesis, and even the cause and genetic factors of the disease. myelodysplastic syndrome (MDS), myeloproliferative neoplasms (myeloproliferative neoplasms, MPN)...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/577G01N33/574G01N33/569G01N33/68G01N33/58G01N15/14
CPCG01N15/14G01N33/56966G01N33/57407G01N33/57492G01N33/577G01N33/582G01N33/68G01N33/6869G01N33/6893G01N2333/70503G01N2333/70507G01N2333/70514G01N2333/70535G01N2333/70539G01N2333/70553G01N2333/70582G01N2333/70589G01N2333/70596G01N2333/7155G01N2800/60
Inventor 王卉陈曼傅旻婧
Owner 信纳克(北京)生化标志物检测医学研究有限责任公司
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