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.
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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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