Bioactive peptide AGYDVEKNNSRIKLGLK and preparation method and application thereof
A bioactive peptide and active technology, applied in the field of protein, can solve problems such as the research of peptide fragments without protein, and achieve the effect of improving the ability, good anti-inflammatory activity, and promoting the increase of induction
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Embodiment 1
[0033] The artificial synthesis of embodiment 1 active peptide AGYDVEKNNSRIKLGLK
[0034] 1. Synthesis of Bioactive Peptides
[0035] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak with 50ml of dichloromethane (DCM).
[0036] After 2.2 hours, the resin was washed with nitrogen-dimethylformamide (DMF) 3 times the volume of the resin, and then drained. This was repeated four times, and the resin was drained for use.
[0037] 3. Add a certain amount of 20% piperidine (piperidine / DMF=1:4, v:v) into the reactor, and shake it on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, wash four times with 3 times the resin volume of DMF, and then drain.
[0038] 4. Take a small amount of resin and test it with ninhydrin (Ninhydrin Nine Wells) method (two drops each of test A and test B, react at 100°C for 1 min), if the resin has color, it means that the deprotection is successful.
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Embodiment 2
[0076] The immune activity experiment of embodiment 2 bioactive peptides
[0077] 1. The experiment of bioactive peptide AGYDVEKNNSRIKLGLK promoting macrophages to secrete cytokines (ELISA method)
[0078] 1. Experimental reagents and instruments:
[0079] Reagents: Experimental animal balb / c mice (male 6, 8 weeks old), Shanghai Slack Experimental Animal Co., Ltd.; mouse lymphocyte extract, Shanghai Suo Laibao Biotechnology Co., Ltd.; RPMI1640 medium, GIBCO Company; cattle Serum albumin (bovine serum albumin, BSA), Genebase Company; The mouse spleen lymphocyte-derived bioactive peptide AGYDVEKNNSRIKLGLK obtained in Example 1; ELISA cytokine rapid kit (TNF-α, IL-1β and IL-6) , Wuhan Boster Biological Engineering Co., Ltd.
[0080] Instruments and equipment: LRH, 250F biochemical incubator Shanghai Heng Technology Co., Ltd.; GL, 22M high-speed refrigerated centrifuge Shanghai Luxiangyi Centrifuge Instrument Co., Ltd.; Hera cell 150 CO2 incubator Heraeus Company; Dragon Wellsca...
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