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Screening method and application of T cells with high anticancer activity

A screening method and anti-cancer activity technology, which is applied in the field of screening high anti-cancer active T cells, can solve the problems of few target cell subgroups, many useless cell subgroups, and low activity, and achieve strong and durable anti-tumor, tumor Long-lasting killing ability and strong re-stimulation amplification effect

Inactive Publication Date: 2018-11-13
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 and application of T cells with high anticancer activity
  • Screening method and application of T cells with high anticancer activity
  • Screening method and application of T cells with high anticancer activity

Examples

Experimental program
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Effect test

Embodiment 1

[0069] A screening method for T cells with high anticancer activity, comprising the following steps:

[0070] (1) Preparation of CD25 antibody-coated magnetic beads:

[0071] 1.1 Take 1mL of particle size of 4.5μm M-450Epoxy Loadable Magnetic Beads (hereinafter referred to as "magnetic beads") (40 million beads / mL), put them on a magnetic stand for 1 min, discard the supernatant, and take out the test tube from the magnetic stand;

[0072] 1.2 Add 1 mL of the first buffer solution (sodium phosphate buffer with pH=8.0) and repeat the washing twice;

[0073] 1.3 Resuspend the magnetic beads in the first buffer solution, add 200 μg of CD25 antibody, control the total volume to 1 mL, and mix well;

[0074] 1.4 Place the resulting mixed solution on a shaker or rotator at 20°C and gently shake, and incubate in the dark for 20 hours;

[0075] 1.5 Then add bovine serum albumin (BSA), control its concentration in the resulting dark incubation solution to be 0.02% (w / v), and then in...

Embodiment 2

[0097] A screening method for T cells with high anticancer activity, comprising the following steps:

[0098] (1) Preparation of CD25 antibody-coated magnetic beads:

[0099] 1.1 Take 1mL of particle size of 10μm M-450Epoxy Loadable Magnetic Beads (hereinafter referred to as "magnetic beads") (40 million beads / mL), put them on a magnetic stand for 1 min, discard the supernatant, and take out the test tube from the magnetic stand;

[0100] 1.2 Add 1 mL of the first buffer solution (sodium borate buffer with pH=8.4) and repeat the washing 3 times;

[0101] 1.3 Resuspend the magnetic beads in the first buffer solution, add 300 μg of CD25 antibody, control the total volume to 1 mL, and mix well;

[0102] 1.4 Place the resulting mixed solution on a shaker or rotator at 18°C ​​and shake gently, and incubate in the dark for 24 hours;

[0103] 1.5 Then add bovine serum albumin (BSA), control its concentration in the resulting dark incubation solution to be 0.02% (w / v), and incubat...

Embodiment 3

[0121] A preparation method of CD25 antibody-coated magnetic beads, comprising the following steps:

[0122] 1.1 Take 1mL of particle size of 50-100μm M-450Epoxy Loadable Magnetic Beads (hereinafter referred to as "magnetic beads") (40 million beads / mL), put them on a magnetic stand for 1 min, discard the supernatant, and take out the test tube from the magnetic stand;

[0123] 1.2 Add 1 mL of the first buffer solution (Tris-hydrochloric acid buffer with pH=9.0) and repeat the washing 3 times;

[0124] 1.3 Resuspend the magnetic beads in the first buffer solution, add 150 μg of CD25 antibody, control the total volume to 1 mL, and mix well;

[0125] 1.4 Place the resulting mixed solution on a shaker or rotator at 35°C and shake gently, and incubate in the dark for 16 hours;

[0126] 1.5 Then add bovine serum albumin (BSA), control its concentration in the resulting dark incubation solution to be 0.02% (w / v), and then incubate at 35°C for 10min;

[0127] 1,6 After the incuba...

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Abstract

The invention provides a screening method of T cells with high anticancer activity, comprising the steps of providing isolated human peripheral blood mononuclear cells; providing CD25 antibody coatedmagnetic beads; detecting the ratio of CD3 and CD25 double-positive cells; using the CD25 antibody coated magnetic beads to screen off CD25 positive cells to obtain a target cell subgroup. The CD25 antibody coated magnetic beads are used herein for the first time to screen the T cell specific subgroup of high anticancer activity; the screening method has good simplicity and operational convenience, is low in cost and helps conveniently attain high-quality cell immune products having good isolate growth condition, great re-stimulated amplification, high tumor-killing capacity and good durability. The invention also provides application of the screening method in immune cell 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 mode and 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 isolate 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 (PBMCs) for culture expansion, and the unsc...

Claims

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Application Information

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IPC IPC(8): C12N5/0783A61P35/00
CPCA61K35/17C12N5/0636C12N2509/00
Inventor 张长风
Owner SHENZHEN BINDEBIOTECH CO LTD
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