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
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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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