Magnetic immune chromatography method capable of synchronously detecting Tm and Pa allergens
A technology of magnetic immunochromatography and simultaneous detection, which is applied in clinical detection of allergens, food inspection, and detection of food allergens. The effect of strong stability
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preparation example Construction
[0069] The preparation method of the magnetic immunochromatography test paper strip which can detect Tm and Pa allergen simultaneously comprises the following steps:
[0070] C. Preparation and purification of antigen: Tm allergen uses shrimp as raw material, Pa allergen uses crucian carp as raw material
[0071] Raw materials were prepared and purified by methods designed by the laboratory.
[0072]D. Preparation and purification of antibodies: First, multiply the specific antibody cell lines of Tm and Pa allergens
[0073] cultured, and then injected into BALB / C mice to prepare ascites; the prepared ascites was concentrated with 50% (w / v) ammonium sulfate and then used commercial protein-G affinity chromatography column, according to the attached product instructions Purify.
[0074] C. Preparation of immunomagnetic beads: select magnetic beads with a diameter of 50-200 nm, and use carbodimethylamide (EDC) and succinimide (NHS) to covalently cross-link anti-Tm monoclonal a...
Embodiment 1
[0091] Embodiment 1: Magnetic immunochromatography detects the main allergen Tm of crustaceans
[0092] (1) Preparation and purification of main crustacean allergen Tm and fish main allergen Pa: take shrimp acetone
[0093] 2g powder, add 30mL 20mmol / L Tris-HCl (pH 7.5) solution containing 1 mol / L KCl to extract overnight, after centrifugation, take the supernatant and put it in a water bath at 100 ℃ for 10min, then centrifuge to recover the supernatant. The resulting supernatant was treated by PI 4.5 isoelectric point precipitation, and the precipitate was dissolved in 20mmol / L Tris-HCl (pH 7.5), adjusted to pH 7.5 with 1 mol / L NaOH, and subjected to 40% ammonium sulfate salting out, and recovered after centrifugation The supernatant was subjected to the isoelectric point operation and salted out with 60% ammonium sulfate. After centrifugation, the precipitate was dissolved in PBS and stored at -20°C for later use.
[0094] The purification method of the main fish allergen P...
Embodiment 2
[0101] Embodiment 2: Magnetic immunochromatography detects fish main allergen Pa
[0102] Except that in the sample detection step, the detection sample is the purified main fish allergen Pa, other steps are the same as in Example 1.
[0103]
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