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T cell activated by bispecific antibody as well as preparation method and application thereof

A bispecific antibody and cell technology, applied in the field of cellular immunology, can solve the problems of strong cell activity, strong expansion ability, and strong cytotoxicity, and achieve the effect of reliable cell quality, simple operation, and controllable technical links

Active Publication Date: 2016-02-03
高岱清 +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in the above scheme, through the connection of bispecific antibodies, tumor cells can present antigens to T cells, then activate T cells, and then kill target cells, so that the prepared tumor cell-specific polyclonal T cells have high specificity. Due to its high specificity, it produces a specific killing effect. Tumor cell-specific polyclonal T cells can only kill target cells, but have no killing effect on non-target cells
[0005] In view of the fact that a method has not been proposed in the prior art to prepare T cells with non-specific killing effect with strong expansion ability, strong cell activity, strong cytotoxicity and large number of cells

Method used

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  • T cell activated by bispecific antibody as well as preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0040] The method for preparing bispecific antibody-activated T cells described in this example specifically includes the following steps:

[0041] S1. Collect 20ml of peripheral blood, centrifuge at 1800rpm for 20 minutes to separate the cells and plasma, take out the plasma and put it in a water bath at 56°C for 30 minutes to inactivate complement, collect the cells and plasma, and set aside;

[0042] S2. Dilute the cells obtained in the step S1 into a cell suspension with 20ml of normal saline, set aside, then get 2 50ml centrifuge tubes, and add 15-20ml of lymphocyte stratification solution (specific gravity 1.077-1.078; Austria) to each test tube Produced by PAA Company), the above-mentioned cell suspension is slowly added to the lymphocyte stratification liquid, and the interface between the two liquids is guaranteed to be clear, then separated at 2700rpm for 10 minutes, then the cell layer of the interface is sucked, and the Wash once with normal saline to obtain mononu...

Embodiment 2

[0051] The method for preparing bispecific antibody-activated T cells described in this example specifically includes the following steps:

[0052] S1. Collect 30ml of peripheral blood, centrifuge at 2200rpm for 5 minutes to separate cells and plasma, take out the plasma and put it in a water bath at 61°C for 3 minutes to inactivate complement, collect the cells and plasma, and set aside;

[0053] S2, dilute the cells obtained in the step S1 into a cell suspension with normal saline, set aside, then get 2 50ml centrifuge tubes, add 15-20ml lymphocyte stratification solution (specific gravity 1.077-1.078; Austrian PAA) to each test tube produced by the company), slowly add the above cell suspension to the lymphocyte stratification liquid, and ensure that the interface between the two liquids is clear, then centrifuge at 2300rpm for 30 minutes, then absorb the cell layer at the interface, and use The mononuclear cells obtained were washed with normal saline for 6 times and used ...

Embodiment 3

[0062] The method for preparing bispecific antibody-activated T cells described in this example specifically includes the following steps:

[0063] S1. Collect 30ml of peripheral blood, centrifuge at 2000rpm for 10 minutes to separate the cells and plasma, take out the plasma and put it in a water bath at 56°C for 35 minutes to inactivate complement, collect the cells and plasma, and set aside;

[0064] S2, dilute the cells obtained in the step S1 into a cell suspension with saline, set aside, then get 2 50ml centrifuge tubes, add 15-20ml lymphocyte stratification solution (specific gravity 1.077-1.078; Austrian PAA company) production), slowly add the above cell suspension to the lymphocyte stratification liquid, and ensure that the interface between the two liquids is clear, then centrifuge at 2500rpm for 20 minutes, then absorb the cell layer at the interface, and use physiological Washed once with saline, the obtained mononuclear cells were set aside;

[0065] S3. Add the...

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Abstract

The invention relates to a T cell activated by a bispecific antibody as well as a preparation method and an application thereof. The preparation method comprises the following steps: S1, taking peripheral blood for centrifugation, separating cells and blood plasma, inactivating complements in blood plasma, collecting cells and blood plasma for standby; S2, diluting the obtained cells to a suspension, separating mononuclear cells, and washing mononuclear cells for standby; S3, adding the obtained mononuclear cells into a serum-free nutrient solution, adding a bispecific antibody, a target cell, a cytokine and blood plasma, carrying out an incubation and cultivation according to a conventional method, collecting cultivated cells, carrying out centrifugal washing with normal saline for obtaining the T cell activated by the bispecific antibody; the bispecific antibody has a function for activating the T cell, the dissociation coefficient between the bispecific antibody and a T cell surface antigen is higher than 1x10<-7>, and the dissociation coefficient between the bispecific antibody and the target cell surface antigen is lower than 1x10<-9>; the molecular weight of the bispecific antibody is less than 50KD; the T cell activated by the bispecific antibody is obtained by the preparation method.

Description

technical field [0001] The invention belongs to the technical field of cellular immunology, and specifically relates to a T cell activated by a bispecific antibody, a preparation method and an application thereof. Background technique [0002] The adoptive immunotherapy of tumors is to inject autologous or allogeneic immunocompetent cells into the patient's body after in vitro expansion, directly kill tumor cells, regulate and enhance the immune function of the body, mainly including LAK cells, CD3AK cells, TIL cells, CTL cells, Immunotherapy of NK cells and CIK cells. As a new method in the comprehensive treatment of tumors, adoptive immunotherapy has been widely combined with conventional surgery, radiotherapy, chemotherapy, cytokines and molecular targeted therapy, and has become an integral part of comprehensive treatment of tumors. It has been shown in the treatment of various tumors broad application prospects. [0003] Among them, T-cell (ie, T lymphocyte) transfusi...

Claims

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Application Information

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IPC IPC(8): C12N5/0783A61K35/17A61K39/395A61P35/00A61P31/00
Inventor 高岱清朱化星杨兆勇高天一孙伟红朱丹妮魏晓芳郭庆明汪康游
Owner 高岱清
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