Screening method for T cells with high antitumor activity and application of screening method

A screening method and anti-cancer activity technology, which is applied in the field of screening T cells with high anti-cancer activity, can solve the problems of many useless cell subsets, poor quality, and few target cell subsets

Inactive Publication Date: 2018-06-05
SHENZHEN BINDEBIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In view of this, the purpose of the present invention is to provide a screening method for T cells with high anti-cancer activity, aiming to solve the problem of many useless cell subsets, few target cell subsets, low activity and poor quality in immune cells in the prior art. question

Method used

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  • Screening method for T cells with high antitumor activity and application of screening method
  • Screening method for T cells with high antitumor activity and application of screening method
  • Screening method for T cells with high antitumor activity and application of screening method

Examples

Experimental program
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Effect test

Embodiment 1

[0067] A method for screening T cells with high anticancer activity, comprising the following steps:

[0068] (1) Preparation of CD27 antibody-coated magnetic beads:

[0069] 1.1 Take 1mL of the particle size of 4.5μm M-450Epoxy loadable magnetic beads (hereinafter referred to as "magnetic beads") (40 million particles / mL), put them on the magnetic stand for 1min, discard the supernatant, and take the test tube out of the magnetic stand;

[0070] 1.2 Add 1 mL of the first buffer solution (sodium phosphate buffer solution with pH=8.0) and wash twice repeatedly;

[0071] 1.3 Resuspend the magnetic beads in the first buffer solution, add 200 μg of CD27 antibody, control the total volume to 1 mL, and mix thoroughly;

[0072] 1.4 Place the obtained mixed solution on a shaker or rotator at 20°C and gently shake it, and incubate in the dark for 20 hours;

[0073] 1.5 Then add bovine serum albumin (BSA), control its concentration in the resulting light-proof incubation solution to...

Embodiment 2

[0095] A method for screening T cells with high anticancer activity, comprising the following steps:

[0096] (1) Preparation of CD27 antibody-coated magnetic beads:

[0097] 1.1 Take 1mL of the particle size of 10μm M-450Epoxy loadable magnetic beads (hereinafter referred to as "magnetic beads") (40 million particles / mL), put them on the magnetic stand for 1min, discard the supernatant, and take the test tube out of the magnetic stand;

[0098] 1.2 Add 1 mL of the first buffer solution (sodium borate buffer solution with pH=8.4) and repeat washing 3 times;

[0099] 1.3 Resuspend the magnetic beads in the first buffer solution, add 300 μg of CD27 antibody, control the total volume to 1 mL, and mix thoroughly;

[0100] 1.4 Place the obtained mixed solution on a shaker or a rotator at 18°C ​​and shake gently, and incubate in the dark for 24 hours;

[0101] 1.5 Then add bovine serum albumin (BSA), control its concentration in the resulting dark incubation solution to 0.02% (w...

Embodiment 3

[0119] A preparation method for CD27 antibody-coated magnetic beads, comprising the following steps:

[0120] 1.1 Take 1mL of the particle size of 50-100μm M-450Epoxy loadable magnetic beads (hereinafter referred to as "magnetic beads") (40 million particles / mL), put them on the magnetic stand for 1min, discard the supernatant, and take the test tube out of the magnetic stand;

[0121] 1.2 Add 1 mL of the first buffer solution (Tris-hydrochloric acid buffer solution with pH=9.0) and repeat washing 3 times;

[0122] 1.3 Resuspend the magnetic beads in the first buffer solution, add 150 μg of CD27 antibody, control the total volume to 1 mL, and mix thoroughly;

[0123] 1.4 Place the obtained mixed solution on a shaker or rotator at 35°C and gently shake, and incubate in the dark for 16 hours;

[0124] 1.5 Then add bovine serum albumin (BSA), control its concentration in the resulting dark incubation solution to 0.02% (w / v), and incubate at 35°C for 10 minutes;

[0125] 1,6 A...

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Abstract

The invention provides a screening method for T cells with high antitumor activity. The screening method comprises the following steps: providing isolated human peripheral blood mononuclear cells; providing CD27 antibody-coated magnetic beads; detecting a ratio of CD3 and CD27 double positive cells; and screening a CD17 positive target cell subset by using the CD27 antibody-coated magnetic beads.The screening method provided by the invention has the advantages that the CD27 antibody-coated magnetic beads are adopted for screening a T cell specific subset with high antitumor activity for the first time, operation is simple and easy, cost is low, and a high-quality cellular immunity product which has good in-vitro growth condition, strong reexcitation proliferation and high and durable tumor killing capability can be conveniently obtained. The invention also provides application of the screening method in immunocyte therapy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a screening method and application of T cells with high anticancer activity. Background technique [0002] Immune cell therapy is an emerging tumor treatment model, which has become a global research hotspot, especially with tumor infiltrating cells (TIL), CAR-T (chimeric antigen receptor T cells) / TCR-T (chimeric T cells) Cell) technology is the main representative and has become the main development direction of global biomedicine. Immune cell therapy is the use of biotechnology and biological agents to separate immune cells from patients for in vitro culture, activation and expansion, and then infuse them back into the patient's body to directly kill or stimulate the body's immune response to kill tumor cells and achieve the purpose of treating tumors. [0003] Existing cellular immunotherapy uses unscreened peripheral blood mononuclear cells (PBMC) for culture and expansion, an...

Claims

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Application Information

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IPC IPC(8): C12N5/0783
CPCC12N5/0636C12N2509/00
Inventor 张长风
Owner SHENZHEN BINDEBIOTECH CO LTD
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