Immunomicrosphere chromatography quick HIT antibody detection test paper
An immune microsphere and antibody detection technology, applied in biological testing, measuring devices, analytical materials, etc., can solve the problems of long test report time, complicated clinical operation, complicated operation, etc., to ensure the validity and quality control, and test results. Clear and accurate results with improved sensitivity
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Embodiment 1
[0049] Such as figure 1 Shown, a kind of immune microsphere chromatography rapid HIT antibody detection test paper preparation method comprises the steps:
[0050] S1. Specific steps for preparation of heparin / PF4 complex:
[0051] 40ul of heparin (1000U / ml), dissolved in PBS buffer of pH 7.4, mixed with 94ul of 1.4mg / ml of platelet factor 4, added 56ul of pure water, incubated at room temperature for 30min and turned into a yellow complex solution, then added 4ul 50% sucrose solution and 4ul of 25% trehalose solution were mixed, and 2ul of 1M TAPS buffer solution of pH 9.0 was added for use.
Embodiment 2
[0053] S2. Preparation of protein A particles
[0054] Specific steps: take 100ml of 10% 0.3um fluorescent microspheres, wash 3 times with PH4.5 50mM MES solution, add 1ml of freshly prepared 10mg / ml EDAC solution (prepared with PH4.5 50mM MES), and incubate at room temperature 30min, then centrifuged at 2000rpm for 5min, then washed with 50mM MES solution of pH 4.5, incubated overnight at room temperature with 1ml of 0.65mg / ml recombinant protein A (PBS), incubated with 10% BSA, 100×Tris-EDTA at room temperature for 1 Hours, centrifuged and washed 3 times with 1ml PBS, stored in 4% sucrose, 0.05% sodium azide solution.
Embodiment 3
[0056] S3, preparation of reaction film
[0057] The compound was sprayed onto a Millipore Hi-Flowmylar backed nitrocellulose membrane (25 mm wide, SHF0900425) at 0.075 μl / mm, spraying at a constant velocity of 45 mm / s to form a test line. At the same time, the reference reagent containing 0.25 mg / mL recombinant protein A was mixed with 1% sucrose / 0.5% trehalose / 10 mM TAPS pH 9.0, and the quality control line was sprayed at 0.075 μl / mm at 45 mm / sec. The films were then baked at 56 °C for one week.
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