Antibody composition and application thereof in screening of post-transplant lymphoproliferative disorders (PTLD)

A technology of lymphocyte proliferation and antibody composition, applied in the biological field, can solve the problems of no FCM detection of peripheral blood early screening means, no standardized plan, and severe symptoms of PTLD patients

Active Publication Date: 2021-03-30
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

The diagnosis of PTLD has always been based on pathology as the gold standard, but there are the following problems: ① PTLD occurs acutely, and some patients have severe symptoms, so it is not convenient to collect lymph nodes for pathology; ② Or some lymph nodes are too small and deep to obtain materials; ③ Pathological test reports The time is relatively long; ④ Follow-up of the condition after surgical resection
[0005] So far, only Washington University School of Medicine in 2004 and the inventor team of this case published an article on the use of FCM to help diagnose PTLD in 2010, but all of them were cases diagnosed by clinical manifestations and other laboratory tests. FCM was used as a technology to help diagnosis. And the program is simple and individualized, and no complete normative program has been proposed, let alone FCM detection of peripheral blood as an early screening method

Method used

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  • Antibody composition and application thereof in screening of post-transplant lymphoproliferative disorders (PTLD)
  • Antibody composition and application thereof in screening of post-transplant lymphoproliferative disorders (PTLD)
  • Antibody composition and application thereof in screening of post-transplant lymphoproliferative disorders (PTLD)

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] The preparation of embodiment 1 reagent

[0063] The antibody combinations used in this example are:

[0064]A1 component is: CD19-Pe-Cy7, CD38-APC, CD20-APC-Cy7, CD56 BV421, CD45 V500, the above five monoclonal antibody reagents are mixed according to volume ratio 3:3:2:3:3 in the first container;

[0065] A2 component is: cytoplasmic kappa-FITC, cytoplasmic lambda-PE, and the two monoclonal antibodies are packed in the second container according to the volume ratio of 2:2;

[0066] The antibodies in this example are commercially available, among which, cytoplasmic kappa-FITC and cytoplasmic lambda-PE are products of Dako Company of Denmark, and other fluorescein directly labeled antibodies are products of Becton Dickinson Company of the United States.

[0067] Optionally add red blood cell lysate and put it in the third container, split the membrane breaking agent A and B liquid into the fourth container and the fifth container, put PBS buffer in the sixth container...

Embodiment 2

[0068] Example 2 Processing of Specimen

[0069] According to the results of cell counting, add heparin or EDTA anticoagulated peripheral blood samples into the flow tube, and ensure that the amount of cells added is about 2×10 6 (1×10 6 -1×10 7 ), with a volume not exceeding 160 μl; first add 3ml PBS and incubate at 37°C for 5 minutes, centrifuge at 300-450g for 5 minutes to remove the supernatant, repeat the incubation and centrifugation once, then add 3ml PBS to wash once (mix well, centrifuge at 300-450g for 5 minutes, Remove the supernatant), then add 14 μl of five membrane monoclonal antibody reagents labeled with different fluorescein (ie, component A1 of Example 1) to the flow tube according to Table 1, mix well with the cell suspension and keep away from room temperature Incubate in light for 15 minutes, add 100 μl membrane disrupting agent A solution, incubate at room temperature in the dark for 5 minutes, add 3ml 1× hemolysin, incubate in the dark for 10 minutes t...

Embodiment 3

[0072] Example 3 Detection of Specimen

[0073] The specimens processed according to the method of Example 2 were tested on a 3-laser 8-color FACS Canto II flow cytometer from Becton Dickinson Company in the United States. After obtaining 500,000 cells per tube, the data was analyzed using diva 2.8 software. Among them, CD45 weak expression (dim) / CD38 strong expression (bright, bri) were selected to select plasma cells, and CD19 / side scatter (side scatter, SSC) and CD20 / SSC antibodies were selected to select B cells .

[0074] The evaluation criteria for monoclonal B cells and plasma cells involved in FCM screening for PTLD are as follows:

[0075] (1) Simultaneous B cell validation using CD19 / SSC and CD20 / SSC.

[0076] (2) Judgment of monoclonal B cells: Under normal circumstances, all B cells of CD19 / SSC and CD20 / SSC, and any part of B cells with different expression intensities of CD19 and CD20, should be polyclonal cells, that is, cytoplasmic kappa / Cytoplasmic lambda i...

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Abstract

The invention provides an antibody composition and an application thereof in screening of post-transplant lymphoproliferative disorders (PTLD). The antibody composition comprises a first group of antibodies and a second group of antibodies, wherein the first group of antibodies comprises an anti-CD19 antibody, an anti-CD38 antibody, an anti-CD20 antibody, an anti-CD56 antibody and an anti-CD45 antibody; and the second group of antibodies comprises an anti-cytoplasm (cytoplasmatic, c) kappa antibody and an anti-cytoplasm lambda antibody. The antibody composition disclosed by the invention can be used for PTLD screening of patients suffering from fever and lymphadenitis after transplantation, can be used for monitoring PTLD in the early stage, and is high in sensitivity, good in specificity,convenient to operate, simple, convenient, economical and high in cost performance.

Description

technical field [0001] The invention relates to an antibody composition and its application in screening lymphocyte proliferative diseases after transplantation, and belongs to the field of biotechnology. Background technique [0002] For a long time, various tumors have been serious diseases that endanger human health and human life. With the gradual aggravation of various problems such as environmental pollution and pressure, as well as the prolongation of population life, more advanced detection methods, the incidence and detection of various tumors The output rate increased significantly. Due to the high degree of malignancy of the tumor, a considerable proportion of patients cannot be relieved by conventional surgery, radiotherapy, and chemotherapy, especially for tumors of the lymphoid hematopoietic system. Hematopoietic stem cell transplantation and various organ transplantation have become the only hope for these patients to survive. At present, 50,000 to 60,000 pat...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574G01N33/577G01N33/58G01N15/14
CPCG01N15/14G01N33/57407G01N33/57492G01N33/57496G01N33/577G01N33/582G01N2333/70503G01N2333/70589G01N2333/70596
Inventor 王卉吴雪英王爱先陈曼宫美维甄军毅
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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