A method for removing glycopeptides in deproteinized extract of calf serum
A calf serum and protein removal technology, applied in the field of chemistry, can solve the problems of no specific glycopeptide, complex polypeptide components, increase the risk of allergic reactions, etc., and achieve the effect of reducing the probability of type 4 allergy
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Embodiment 1
[0019] The implementation method of embodiment 1 calf serum deproteinized injection except glycopeptide
[0020] 1. According to the method of Chinese patent application 201510145515.5, extract 1000ml of deproteinized calf serum injection;
[0021] 2. Take 5 parts of the above calf serum deproteinized injection, each 100g, mark sample 1-sample 5 respectively; lower the temperature of the 5 samples to -7°C;
[0022] 3. Add 0.05g, 0.1g, 0.15g, 0.2g, 0.25g of ZnS powder passed through a 200-mesh sieve to samples 1-5 after cooling, and stir evenly;
[0023] 4. Put sample 1-sample 5 into -40°C environment for quick freezing for 4 hours;
[0024] 5. Put the quick-frozen samples 1 to 5 in an environment of -23°C for 6 hours;
[0025] 6. Put sample 1-sample 5 at room temperature to melt slowly;
[0026] 7. Filter each sample to remove ZnS powder.
Embodiment 2
[0027] Embodiment 2 The method of the present invention removes the effect of glycopeptide
[0028] RBL-2H3 cell model was used to detect the effect of removing glycopeptides; the content of glycopeptides as allergens was judged by the degranulation rate of cells.
[0029] Take RBL-2H3 cells in the logarithmic phase and press 2~3×10 5 Each well was inoculated in a 96-well plate with an inoculation volume of 100 μl, and 5 groups were set up, with 5 duplicate wells in each group. After culturing for 24 hours, 10 μl each of sample 1-sample 5 were added to the 5 wells of each group, and shaken gently to mix. Evenly mix the drug, and after 0.5h of co-action between the drug and the cells at 37°C, discard the culture medium, add 200 μl of 0.5% neutral red staining solution to each well, and stain for 3 minutes at 37°C. Count the number of degranulated cells in 100 cells, and calculate the percentage of degranulated cells (average of 5 duplicate wells). The results are shown in Tab...
Embodiment 3
[0034] Embodiment 3 The method of the present invention affects the quality of calf serum deproteinized injection
[0035] The two core indicators of calf serum deproteinized injection: peptide content and trace respiratory activity stimulation index were selected for detection to judge the impact of this method on product quality. The detection method of the polypeptide content is detected by the lowry protein method, and the trace respiratory stimulation index is detected by the trace breath test method. Take the untreated calf serum deproteinized injection in Example 1 as a blank control. The test results are shown in Table 2.
[0036] The detection result of two indexes before and after the method of the present invention is processed in table 2
[0037] group Peptide content (mg / ml) Trace Respiratory Stimulation Index (SI) sample 1 7.56 11.6 sample 2 7.43 11.7 sample 3 7.31 11.6 Sample 4 7.20 11.5 Sample 5 7.16 11.5 c...
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