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Ten-color antibody composition and application thereof in leukemia-lymphomas subtype

An antibody composition and a technology for the composition, which are applied in the fields of biological material analysis, disease diagnosis, analysis materials, etc., can solve problems such as detection of leukemia/lymphoma that have not yet been found, and achieve the effects of cost saving and simple operation.

Active Publication Date: 2017-05-31
ZHEJIANG BOZHEN BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no leukemia / lymphoma detection kit with a "single-tube ten-color" combination (ten antibody combinations)

Method used

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  • Ten-color antibody composition and application thereof in leukemia-lymphomas subtype
  • Ten-color antibody composition and application thereof in leukemia-lymphomas subtype
  • Ten-color antibody composition and application thereof in leukemia-lymphomas subtype

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0044] The preparation of embodiment 1 reagent

[0045] The antibody combinations used in this example are as follows (each antibody is named after the antigen and the fluorescent label ligature): CD71-FITC, CD7-PE, CD13-ECD, CD33-PerCP-cy5.5 (abbreviated as CD33 PC5.5) , CD19-PerCP-cy7 (abbreviated as CD19PC7), CD117-APC, CD34-APC-Alexa Fluor700 (abbreviated as CD34 APC-A700), CD10-APC-Alexa Fluor750 (abbreviated as CD10 APC-A750), HLA-DR- BV421, CD45-KromeOrange (abbreviated as CD45KO), these antibodies can be purchased directly through public channels, and the antibodies in the examples of the present invention are all purchased from Shanghai Dakowei Biotechnology Co., Ltd.

[0046] The above monoclonal antibodies were sampled, serially diluted, tested on the flow cytometer respectively, and the optimal dosage of each antibody was determined by titration. The specific dosage is shown in the table below.

[0047]

[0048] The above ten kinds of quantitative antibody reag...

Embodiment 2

[0049] The processing of embodiment 2 sample

[0050] Take heparin or EDTA anticoagulated bone marrow or peripheral blood samples, adjust to a cell density of 1-5×10 6 / ml, and then filtered through a 200-mesh industrial sieve to remove agglomerate-like substances in the sample to ensure a single cell suspension state, and store at 2-8°C, that is, the processed specimen.

Embodiment 3

[0051] The detection of embodiment 3 sample

[0052] Take the flow tube and add the antibody mixture in the kit of Example 1. Then take 50 μl of the specimen treated in Example 2, vortex the shaker to mix well, and incubate at room temperature (25° C.) in the dark for 15 minutes.

[0053] Dilute 10× red blood cell lysate into 1× red blood cell lysate with 1× PBS buffer, then add 2ml of the diluted 1× red blood cell lysate to the above-mentioned flow tube after incubation, shake and mix with a vortex shaker, and keep at room temperature (25°C) and stand in the dark for 10 minutes.

[0054] Centrifuge the flow tube at 1500 rpm for 5 minutes, discard the supernatant, and add 200 μl 1×PBS to resuspend. After the sample is processed, it is detected by a flow cytometer, and detected by fluorescently labeled ten-color wavelengths.

[0055] A large number of samples of known hematological tumors from different sources were tested, and CD45 vs SSC was used as a gate to examine the d...

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Abstract

The invention belongs to the technical field of antibodies, and provides an antibody composition composed of ten antibodies, wherein the tan antibodies comprises CD71 resistant antibody, CD7 resistant antibody, CD13 resistant antibody, CD33 resistant antibody, CD19 resistant antibody, CD117 resistant antibody, CD34 resistant antibody, CD10 resistant antibody, HLA-DR resistant antibody, and CD45 resistant antibody. The invention provides a leukemia-lymphomas primary screening kit containing the antibodies, and the application thereof.

Description

technical field [0001] The invention belongs to the technical field of antibodies, in particular, the invention relates to a ten-color antibody composition, its application in leukemia and lymphoma typing, a typing kit and the like. Background technique [0002] Hematological tumors such as leukemia and lymphoma have become one of the most threatening diseases to human health in recent years. The immunophenotyping of leukemia plays a very important role in the diagnosis of the disease. Immunotyping of leukemia, lymphoma, etc. by flow cytometry is one of the important methods for the diagnosis of blood tumors. Our company's patent CN 105606797 B discloses a four-color flow cytometry antibody composition and its application in the classification of leukemia and lymphoma, which contains 7 kinds of fluorescently labeled monoclonal antibodies. A more comprehensive detection of leukemia was performed, but two loadings were required. Patent CN 103675277 A discloses a fluorescent...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/577G01N33/533
CPCG01N33/533G01N33/574G01N33/577G01N2800/7028
Inventor 倪万茂
Owner ZHEJIANG BOZHEN BIOTECH CO LTD
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