Applications of isotope labeled anti-MHCII monoclonal antibody
An isotope labeling, monoclonal antibody technology, applied in instruments, measuring devices, scientific instruments, etc., can solve problems such as damage to donor organs, high price, uneven emission lines, etc., to achieve reduced stimulation, strong feasibility, and strong practicality Effects of Sexuality and Effectiveness
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Embodiment 1
[0020] Embodiment one preparation 188 Re-2E9 / 13F(ab) 2 :
[0021](1) Extract F(ab) from 2E9 / 13 monoclonal antibody after papain hydrolysis 2 Fragments, filtered, purified, concentrated, and stored in a -20°C refrigerator;
[0022] (2) from 188 W- 188 Obtained from the Re generator 188 ReO4 2- eluent, according to the direct preparation method 188 Re is labeled with F(ab) 2 fragments, prepared as 188 Re-2E9 / 13F(ab) 2 fragment.
Embodiment 2
[0023] Example 2 Construction of in vitro unidirectional mixed lymphocyte culture (MLR) model
[0024] ① Set positive control group A (stimulator cells + responder cells after normal saline treatment), monoclonal antibody group B (stimulator cells + responder cells after monoclonal antibody treatment), and isotope-labeled monoclonal antibody group C (stimulator cells after Stimulator cells+response cells) and negative control group D (response cells), each group was set to 48, 72, 96, 120 hours (H) different reaction time, after harvesting cells, do MTT test;
[0025] ② The groups were grouped as above, with different reaction times of 72 and 120 hours for each group, and RT-PCR was performed after the cells were harvested, and comparative analysis was performed.
Embodiment 3
[0026] Example 3 Application in In Vitro Cellular Immune Rejection Model
[0027] ① Separation of mononuclear cells from peripheral blood:
[0028] 1. Take 20ml of peripheral venous blood from donor and recipient pigs (pigs of different strains) respectively, and dilute the peripheral blood by 1:1 with physiological saline.
[0029] II: Take a new sterile centrifuge tube, add FICOLL according to the ratio of diluted peripheral blood and FICOLL at a ratio of 1.2:1, and slowly add peripheral blood at a place 1cm above the FICOLL solution.
[0030] III: Centrifuge (2000-2500 rpm) for 25 minutes
[0031] IV: Draw the mononuclear cell layer between the plasma and FICOLL to another test tube, add PBS solution and blow evenly, add red blood cell lysate, remove excess red blood cells, add more than twice the volume of normal saline, and centrifuge (1000 rpm )10 minutes. Add PBS to wash the lymphocytes three times.
[0032] V: Discard the supernatant, resuspend with 1640 culture me...
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