Applications of isotope labeled anti-MHCII monoclonal antibody

An isotope labeling, monoclonal antibody technology, applied in instruments, measuring devices, scientific instruments, etc., can solve problems such as damage to donor organs, high price, uneven emission lines, etc., to achieve reduced stimulation, strong feasibility, and strong practicality Effects of Sexuality and Effectiveness

Inactive Publication Date: 2008-05-21
ZHEJIANG UNIV
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  • Abstract
  • Description
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AI Technical Summary

Problems solved by technology

[0003] At present, the number of patients receiving organ transplantation is increasing year by year, but the incidence of acute rejection of organs after transplantation is very high, which affects the success rate of transplantation and the survival rate of patients
The existing main methods of anti-immune rejection still rely on various immunosuppressants, which are not only expensive, but also significantly increase the chances of infection and tumor occurrence due to immunosuppression
In addition, the method of reducing the immunogenicity of grafts and prolonging the survival time of transplanted organs by relying on in vitro culture, freezing, external radiation therapy, animal adoptive transplantation and other means has achieved certain results, but it has no practical application in organ transplantation Value, for example, external radiation therapy makes the radiation received inside and outside the donor organ often uneven, the dose is too small to achieve the desired effect, and the dose is too large may cause extensive damage to the donor organ
Therefore, so far, no clinically applicable and effective means have been found.

Method used

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  • Applications of isotope labeled anti-MHCII monoclonal antibody

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment one preparation 188 Re-2E9 / 13F(ab) 2 :

[0021](1) Extract F(ab) from 2E9 / 13 monoclonal antibody after papain hydrolysis 2 Fragments, filtered, purified, concentrated, and stored in a -20°C refrigerator;

[0022] (2) from 188 W- 188 Obtained from the Re generator 188 ReO4 2- eluent, according to the direct preparation method 188 Re is labeled with F(ab) 2 fragments, prepared as 188 Re-2E9 / 13F(ab) 2 fragment.

Embodiment 2

[0023] Example 2 Construction of in vitro unidirectional mixed lymphocyte culture (MLR) model

[0024] ① Set positive control group A (stimulator cells + responder cells after normal saline treatment), monoclonal antibody group B (stimulator cells + responder cells after monoclonal antibody treatment), and isotope-labeled monoclonal antibody group C (stimulator cells after Stimulator cells+response cells) and negative control group D (response cells), each group was set to 48, 72, 96, 120 hours (H) different reaction time, after harvesting cells, do MTT test;

[0025] ② The groups were grouped as above, with different reaction times of 72 and 120 hours for each group, and RT-PCR was performed after the cells were harvested, and comparative analysis was performed.

Embodiment 3

[0026] Example 3 Application in In Vitro Cellular Immune Rejection Model

[0027] ① Separation of mononuclear cells from peripheral blood:

[0028] 1. Take 20ml of peripheral venous blood from donor and recipient pigs (pigs of different strains) respectively, and dilute the peripheral blood by 1:1 with physiological saline.

[0029] II: Take a new sterile centrifuge tube, add FICOLL according to the ratio of diluted peripheral blood and FICOLL at a ratio of 1.2:1, and slowly add peripheral blood at a place 1cm above the FICOLL solution.

[0030] III: Centrifuge (2000-2500 rpm) for 25 minutes

[0031] IV: Draw the mononuclear cell layer between the plasma and FICOLL to another test tube, add PBS solution and blow evenly, add red blood cell lysate, remove excess red blood cells, add more than twice the volume of normal saline, and centrifuge (1000 rpm )10 minutes. Add PBS to wash the lymphocytes three times.

[0032] V: Discard the supernatant, resuspend with 1640 culture me...

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Abstract

The present invention provides the application of isotope- labeled monoclonal antibody. An <188>Re is labeled on a 2E9/13 monoclonal antibody and is dialyzed, depurated, concentrated and filtrated to prepare an <188>Re-2E9/13F (ab) 2. An in vitro unidirectional mixed lymphocyte reaction model is constructed. The antibody is applied to the model to detect the repulsion and the inhibitory actions of the antibody to cellular immunity. The present invention changes the method of applying a single method to construct in the prior art, combines the two methods of biology and nuclear medicine, brings in the monoclonal antibody 2E9/13F (ab) 2 segment of anti-MHC-II molecule labeled by the <188>Re in a repulsion and inhibitory model constructed by a same different-source lymphocyte, and the problem that the immunogenicity of a homologous MHCII like antigen is permanently reduced is solved. The present invention is of reasonable design, practical and valid; and provides strong practical support and theoretical basis to animal in vitro experiments and the final application on clinic.

Description

technical field [0001] The present invention relates to a kind of isotopic 188 Application of Re-labeled anti-MHC-II monoclonal antibody in in vitro unidirectional mixed lymphocyte culture model. Background technique [0002] It is one of the important means of modern medical treatment to replace non-functioning tissues, cells and organs through the transplantation of tissues, cells or organs. The individual who provides the graft is called the donor, and the individual who receives the graft is called the recipient. According to the combination of donor and recipient, there are three types of transplantation: Autologous transplantation refers to the transplantation of tissues or organs from the transplant recipient itself to the recipient. Allogeneic transplantation refers to the transplantation between individuals with identical or very similar genetic structures. Allogeneic transplantation refers to the transplantation between individuals with different genetic structu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/566G01N33/554
Inventor 曹利平刘杰倪俊
Owner ZHEJIANG UNIV
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