Method for detecting humoral immunogenicity of autologous CART cells

An immunogenic, cell-based technology, applied in the field of biomedicine, can solve problems such as inability to repeat treatment again, failure of CART cell therapy, etc., and achieve the effect of avoiding the failure of CART treatment

Pending Publication Date: 2021-07-13
苏州才博医学科技有限公司
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Problems solved by technology

Whether it is pre-existing before the patient receives CART cell therapy, or the immune response against the chimeric antigen receptor CAR induc

Method used

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  • Method for detecting humoral immunogenicity of autologous CART cells
  • Method for detecting humoral immunogenicity of autologous CART cells

Examples

Experimental program
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Embodiment

[0021] Example. Detection method of anti-CAR antibody

[0022] combine figure 1 , including the following steps:

[0023] 1. Take 3-5 ml of anticoagulated peripheral blood from the patient, and use magnetic bead sorting to separate and purify peripheral blood T cells.

[0024] 2. Count no less than 50,000 purified peripheral blood T cells, as negative control target cells, wash 3 times with RPMI culture medium, and finally suspend T cells in 50 microliters of RPMI cell culture medium, place the centrifuge tube in on ice.

[0025] 3. Count the same number of CD19-specific autologous CART cells as the target cells for detecting anti-CAR antibodies, wash 3 times with RPMI culture medium, and finally suspend the CART cells in 50 microliters of RPMI cell culture medium, and place the centrifuge tube Keep on ice.

[0026] 4. Add the same amount of patient serum, not less than 10 microliters, to the centrifuge tubes of the above-mentioned T cells and CART cells.

[0027] 5. Mix ...

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Abstract

The invention provides a method for detecting the humoral immunogenicity of autologous CART cells. According to the invention, autologous T cells, which are not modified by CAR, of a testee are used as negative control target cells, autologous CART cells, which are modified by CAR, of the testee are used as target cells for detection, and the reactivity difference between the detected serum sample of the testee and the two target cells is determined through comparison through a flow cytometry, so that whether the anti-CAR antibody exists in the detected serum sample of the testee or not is judged; and the method not only can help a clinician to screen a CAR transformation method which is not easy to cause generation of an anti-CAR antibody for a patient before CART treatment is carried out on the patient, but also can be used as an etiological detection means for dynamically monitoring CART treatment failure possibly caused by induction of the anti-CAR antibody in CART cell treatment, so that CART cells play the maximum treatment effect.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for detecting humoral immunogenicity of autologous CART cells. Background technique [0002] Chimeric Antigen Receptor T cell (CART, Chimeric Antigen Receptor T cell) is a receptor (CAR) that can recognize the specific target of tumor cells, which is chimerically expressed on the surface of T cells of patients through genetic engineering methods, so that the CAR can The modified autologous T cells can recognize and kill tumor cells in the patient's body for the treatment of tumors. [0003] Clinical experiments have found that the patient's autologous T cells have been genetically engineered to embed a chimeric antigen receptor (CAR) on the surface of the T cells that can specifically bind to the B cell surface marker CD19. This modified autologous T cell, That is, CD19-CART cells have achieved very significant curative effects in the treatment of certain malignant tumors of ...

Claims

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Application Information

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IPC IPC(8): G01N33/68G01N33/569G01N33/58
CPCG01N33/56966G01N33/582G01N33/6854G01N2469/20
Inventor 蔡俊超
Owner 苏州才博医学科技有限公司
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