Group of markers for predicting storm of immune checkpoint of COVID-19, application and kit thereof
An immune checkpoint, coronavirus technology, applied in the field of biomedicine, can solve problems such as failure to predict patient biomarkers, decreased immune response levels, and decreased antiviral ability, achieving the effects of rapid detection, broad application prospects and low cost
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[0032] In a specific embodiment of the present invention, the preparation method of the kit preferably includes the following steps:
[0033] (1) Will be coated with one or two of BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 The above capture antibodies were coupled to the encoded microspheres, and the resulting microspheres were coated with BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 Encoded microspheres of one or more than two capture antibodies;
[0034] (2) One or more detection antibodies in BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 Connect biotin to get one of BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 respectively labeled with biotin One or more detection antibodies;
[0035] There is no sequence relationship between the steps (1) and (2).
[0036] The present invention combines one or more capture antibodi...
Embodiment 1
[0061] Preparation of liquid chip kits for detection of BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 biomarkers.
[0062] 1. Kit Composition
[0063] (1) 13-plex coated microspheres: containing BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 capture antibody-encoded microspheres;
[0064] (2) 13-plex biotin-labeled detection antibodies: BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 detection antibodies;
[0065] (3) streptavidin phycoerythrin.
[0066] Among them, the clone numbers of capture antibodies BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27 and CD152 are 6F7, 110416, LH1, The clone numbers of 7H8L17, C9B7W, J116, MIH1, F38-2E2, 10F3, 18, 4B4, O323, 14D3; BTLA, GITR, HVEM, IDO, PD-L1, CD28, CD80, and CD152 detection antibodies are MIH26, DT5D3, eBioHVEM-122, 2E2.6, 10F.9G2, 37407, MEM-233, WKH 203, the detection antibodies of ...
Embodiment 2
[0088] BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27, and CD152 Liquid Chip Kits in Predicting COVID-19 Immune Checkpoint Storm Applications.
[0089] 1. Purpose of the experiment
[0090] Demonstrated that baseline concentrations of BTLA, GITR, HVEM, IDO, LAG-3, PD-1, PD-L1, TIM-3, CD28, CD80, 4-1BB, CD27, and CD152 were significantly more 19 patients were moderately high.
[0091] 2. Subjects
[0092] 1) On the premise of informed consent and meeting the inclusion conditions, select the enrolled cases and record personal basic information. The cohort of COVID-19 patients came from 109 COVID patients in Beijing Ditan Hospital affiliated to Capital Medical University, including 5 asymptomatic patients, 60 mild or moderate patients, and 44 severe or critical patients.
[0093] 2) Plasma was drawn from the patients before treatment when they were admitted to the hospital, and the samples were stored in a -80°C refrigerator.
[0094] 3. Reagent Pr...
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