Fraxinus pennsylvanica pollen allergen extract and immersion liquid of fraxinus pennsylvanica pollen allergen extract and preparation method thereof
A technology for allergens and extracts, which is applied in the preparation methods of peptides, allergen antigen components, chemical instruments and methods, etc., can solve the problems of uncertain components, decreased positive rate, poor stability, etc., and achieves short cycle times. , quick effect, high specificity effect
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Embodiment 1
[0067] The identification of embodiment 1 white wax pollen
[0068] Since the identification and purity of raw materials are extremely important for subsequent allergen diagnosis and therapeutic preparations, this example uses DNA-specific sequence detection and microscopic examination to perform double identification and quality control on the raw material of white wax.
[0069] 1. DNA extraction from white wax pollen
[0070] Tiangen Rapid DNA Extraction and Amplification Kit (Tiangen Biochemical KG203) was used.
[0071] Extraction method: Weigh 5mg of wax pollen sample and place it in a 1.5mL centrifuge tube, add 100μl of Buffer 1, grind the sample thoroughly with a disposable grinding pestle, add 100μl of Buffer 2, shake and mix, centrifuge at 12000r / min for 5min Then take the supernatant and put it in a new centrifuge tube as a DNA template for later use.
[0072] 2. Primer design and synthesis
[0073] The primer sequences are as follows:
[0074] Fra ITS2-F (SEQ ID...
Embodiment 2
[0089] Example 2 Degreasing and Extracting Key Process Steps of Important Parameters of Pyrex Pollen
[0090] 1. Degreasing process parameters of white wax pollen degreasing process
[0091](1) Degrease treatment with the white wax pollen (g) and acetone (ml) at a weight-to-volume ratio (w / v) of 1:2. The fat content of samples of different batches (different collection time) of white wax pollen with different degreasing time was tested to determine the optimal degreasing time.
[0092] Haineng SOX500 fat measuring instrument was used to measure the fat content in the pollen of white wax pollen. Through the method of Soxhlet extraction and dry weighing, the change of pollen weight before and after extraction was compared to obtain the corresponding fat content, and the fat content was compared after degreasing. Pollen for quality control.
[0093] see results figure 2 , it can be seen that after the acetone degreasing time is 30min, the fat content hardly decreases with the...
Embodiment 3
[0104] Embodiment 3 white wax pollen raw material technology
[0105] 1. Pollen collection
[0106] Natural shedding method to collect white wax pollen. Normal temperature or vacuum or fluidized bed drying, until the pollen no longer sticks. Pass the dried pollen through a 150-250 mesh sieve, preferably a 180 mesh sieve in this embodiment.
[0107] 2. Dry
[0108] Spread the pollen in a ventilated and dry place to dry naturally, or vacuum dry, or dry in a fluidized bed for 6-48 hours, until the pollen no longer adheres.
[0109] 3. Degreasing
[0110] The pollen (g) obtained above and acetone (ml) are subjected to degreasing treatment at a weight-to-volume ratio (w / v) of 1:5-1:1. In this embodiment, it is preferably 1:2. Stir or oscillate to degrease for 30 minutes, let stand to separate layers, and observe the clarification of the upper liquid. Pour out the upper layer, add new acetone, and repeat degreasing until the upper layer is clear.
[0111] 4. Dry again
[01...
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