Bioactive peptide with amino acid structure VVRKPLNKEGKKP as well as preparation method and application of bioactive peptide
A bioactive peptide and amino acid technology, applied in the field of protein
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Embodiment 1
[0034] The artificial synthesis of embodiment 1 active peptide VVRKPLNKEGKKP and VVRKPLNKEGKKPR
[0035] 1. Synthesis of Bioactive Peptides
[0036] 1. Weigh 3g of RINK resin (degree of substitution: 0.3mmol / g) into a 150ml reactor and soak with 50ml of dichloromethane (DCM).
[0037] 2. After 2 hours, wash the resin with nitrogen-dimethylformamide (DMF) that is 3 times the volume of the resin, and then drain it. Repeat this four times, and drain the resin for use.
[0038] 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.
[0039] 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...
Embodiment 2
[0078] Example 2 Immunomodulatory Activity Experiment of Bioactive Peptides
[0079] 1. Determination of the amount of bioactive peptide VVRKPLNKEGKKP that promotes nitric oxide induction in macrophages (Griess method)
[0080] 1. Experimental reagents and instruments:
[0081]Reagents: experimental animal balb / c mice (male 6-8 weeks old) spleen lymphocyte-derived bioactive peptide VVRKPLNKEGKKP; LPS, purchased from Sigma Company; neutral red staining solution, produced by Beyond Biotechnology Research Institute.
[0082] Equipment: LRH-250F biochemical incubator Shanghai Heng Technology Co., Ltd.; GL-22M high-speed refrigerated centrifuge Shanghai Luxiangyi Centrifuge Instrument Co., Ltd.; Hera cell 150CO 2 Incubator Heraeus; Dragon WellscanMK3 microplate reader Labsystems.
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