Immune quantitative detection device for marker to be detected, detection method and application
A technology of immunological quantification and detection method, which is applied in the direction of measuring devices, biological testing, material inspection products, etc., and can solve the problems that immune detection technology cannot be realized at the same time
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Embodiment 1
[0081] Embodiment 1: the preparation of filter of the present invention
[0082] Experimental materials: commercially available microfilters, NHS activated agarose gel particles (ACRO BIOSYSTEMS), anti-human fibrinogen polyclonal antibody (Genagates Inc.), sodium bicarbonate, hydrochloric acid, ethanolamine.
[0083] Experimental method: Take a commercially available microfilter and take out the filter element; take 5 mg of anti-human fibrinogen polyclonal antibody plus 0.2M sodium bicarbonate solution, pH 8.3, and dissolve in 2ml; take 1ml of NHS-activated agarose gel particles, and use 20ml of 1mM hydrochloric acid, wash by suction filtration three times; mix the anti-human fibrinogen polyclonal antibody solution with the treated NHS-activated agarose gel particles, and shake for 4 hours at 4°C; the NHS-activated agarose gel after washing reaction Then add 10mM ethanolamine, 0.2M sodium carbonate solution, pH 8.0, shake at room temperature for 4 hours to block the uncoupled ...
Embodiment 2
[0084] Embodiment 2: Comparison of detection performance between the present invention and existing chemiluminescence detection technology
[0085] Experimental materials: anti-human fibrinogen polyclonal antibody filter, horseradish peroxidase-labeled anti-human fibrinogen monoclonal antibody, magnetic particles, luminol, p-iodophenol, carbamide peroxide, chemiluminescence detector, human Fibrinogen solution.
[0086] Experimental method: Take human fibrinogen solution of known concentration, dilute it with PBS solution to prepare 1ug / ml human fibrinogen solution. The experimental method will use the present invention and the current chemiluminescence detection technology to observe the influence of different binding reaction times on the amount of luminescence. Observe the binding reaction time points of 1 minute, 2 minutes, 4 minutes, 10 minutes, 20 minutes, 30 minutes, 45 minutes, and 60 minutes.
[0087] For the current chemiluminescence detection technology group, add ...
Embodiment 3
[0090] Embodiment 3: Comparison of the present invention with the detection results of the current chemiluminescence detection technology
[0091] Experimental materials: anti-human fibrinogen polyclonal antibody filter, horseradish peroxidase-labeled anti-human fibrinogen monoclonal antibody, magnetic particles, luminol, p-iodophenol, carbamide peroxide, chemiluminescence detector, human Fibrinogen solution, healthy human plasma.
[0092] Experimental method: Take a human fibrinogen standard solution of known concentration, dilute it with PBS solution to prepare 10, 30, 70, 100, 300, 700ng / ml human fibrinogen solution; take another healthy person's plasma, and use PBS for 10000 times Dilution; experimental method adopts the present invention and existing chemiluminescence detection technology to detect human fibrinogen solution and draws standard curve, then measures healthy human fibrinogen, and calculates fibrinogen concentration with standard curve; Get 42 test tubes, be d...
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