An antibody chip kit for biomarker screening
A technology of biomarkers and antibody chips, applied in the field of biomedicine, can solve the problems of low sensitivity, cumbersome operation, expensive instruments, etc., achieve high-throughput screening, overcome cumbersome operations, and reduce the amount of specimens.
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Embodiment 1
[0018] Example 1: Preparation of an antibody chip kit for screening biomarkers.
[0019] In order to screen the corresponding biomarkers in patient specimens, the base membrane immobilized with specific antibodies against the following proteins: angiopoietin, brain-derived neurotrophic factor, B-lymphocyte chemotactic factor, bone morphogenetic protein- 4. Bone morphogenetic protein-6, CKβ8-1, ciliary neurotrophic factor, epidermal growth factor, chemokine, chemokine-2, chemokine-3, fibroblast growth factor-6, fibroblast Growth factor-7, FMS-like tyrosine kinase 3 ligand, chemokine CX3CL1, chemokine (C-C motif) ligand 6, glial cell-derived neurotrophic factor, granulocyte colony-stimulating factor, chemokines (C-C motif) ligand 1, γ-interferon, insulin-like growth factor binding protein-1, insulin-like growth factor binding protein-2, insulin-like growth factor binding protein-4, insulin-like growth factor 1, interleukin -10, Interleukin-13, Interleukin-15, Interleukin-16, In...
Embodiment 2
[0033] Embodiment 2: Experiment of detecting cytokines with the kit of the present invention.
[0034] Put the bottom film in the supporting square box, since each bottom film in this embodiment is distributed with a chip lattice, so in this embodiment, the square box is provided with 8 squares, and each square is placed with a bottom film , each chip dot matrix becomes a mutually independent reaction area through the grid. The box with 8 grids used in this example was self-made by the inventor. After adding 2 ml of blocking solution to each grid, place it at room temperature and incubate for 30 minutes, and then perform the following steps in sequence:
[0035] 1. Add sample
[0036] Aspirate the blocking solution in each square, put 1-2 ml of the sample diluted with the blocking solution into the square with a membrane, and then shake it on a shaker at room temperature for 1 to 2 hours, or at 4°C Under reaction 12-18 hours.
[0037] 2. Membrane washing
[0038] Washing s...
Embodiment 3
[0053] Example 3: Test of cross-reactivity of the kit of the present invention.
[0054] The cross-reactivity test between antibody pairs was carried out according to the following method: adding the same 60 kinds of cytokine standard proteins to different bottom membranes. After incubation, slides are washed and then incubated with individual detection antibodies for each antigen. Finally, it was incubated with horseradish peroxidase-labeled streptavidin, and the chip was scanned and read. Taking the capture antibody of each antigen as the horizontal axis and the added detection antibody as the vertical axis, the experimental results in Table 2 can be obtained.
[0055] Table 2 Cross-reaction test results between antibody pairs
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[0072] IL-13
[0073] The experimental ...
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