A kind of identification method and application of bee pollen allergenic protein
An identification method and technology of allergenic proteins, which can be used in instruments, measuring devices, scientific instruments, etc., and can solve problems such as imperfect identification of allergenic proteins.
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Embodiment 1
[0059] This embodiment provides a method for identifying rape bee pollen allergens, and the specific steps are as follows:
[0060] 1) Sample pretreatment to obtain bee pollen freeze-dried powder:
[0061] Accurately weigh 5.0g of ground rapeseed bee pollen powder (100-300 mesh);
[0062] The bee pollen powder is subjected to vacuum freeze-drying by using a vacuum freeze-drying machine, and the required drying time is about 48 hours to obtain the bee pollen freeze-dried powder.
[0063] Use GB5009.3-2016 "National Food Safety Standard for Determination of Moisture in Food" to measure the moisture content of the freeze-dried powder, which meets the standard below 7%.
[0064] 2) Protein extraction to obtain precipitated protein:
[0065] Accurately weigh 100.0 mg rape bee pollen freeze-dried powder, add 1 mL protein extractant;
[0066] Ultrasonic crushing, the condition is ultrasonic 3s, interval 3s, repeat 30 times, continue to shake at room temperature for 30~60min after ...
Embodiment 2
[0099] This example provides an identification method for known allergenic proteins in rapeseed bee pollen to verify the accuracy of the identification method described in the present invention.
[0100] The specific experimental steps are the same as in Example 1. After identification, a known allergenic protein in rape bee pollen is Polcalcin. At the same time, the primary sequence of Polcalcin is obtained, and the T cells and T cells of the allergenic protein are obtained through structural analysis. B cell antigen epitopes were positive, consistent with the characteristics of known allergenic proteins.
[0101] It can be seen that the above test results have verified that the identification method of the present invention is accurate and reliable.
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