A kind of bioactive peptide saprhgslgflprk and its preparation method and application
A technology of biologically active peptides and drugs, applied in the protein field, to achieve the effects of promoting in vitro proliferation, good anti-inflammatory activity, and improving the ability of
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Embodiment 1
[0033]Example 1 Artificial synthesis of active peptide SAPRHGSLGFLPRK
[0034] 1. Synthesis of Bioactive Peptides
[0035] 1. Weigh 3 g of RINK resin (degree of substitution: 0.3 mmol / g) into a 150 ml reactor, and soak it with 50 ml of dichloromethane (DCM).
[0036] After 2.2 hours, the resin was washed with nitrogen-dimethylformamide (DMF) 3 times the resin volume, and then drained. This was repeated four times, and the resin was drained for use.
[0037] 3. A certain amount of 20% piperidine (piperidine / DMF=1:4, v:v) was added to the reactor, and it was shaken on a decolorizing shaker for 20 minutes to remove the Fmoc protecting group on the resin. After deprotection, it was washed four times with 3 times the resin volume of DMF, and then drained.
[0038] 4. Take a small amount of resin and test it by ninhydrin (Ninhydrin hydrate) method (two drops each of test A and test B, react at 100°C for 1 min), the resin is colored, indicating that the deprotection is successful. ...
Embodiment 2
[0075] Example 2 Immunological Activity Experiment of Bioactive Peptides
[0076] 1. MTT assay to determine the in vitro macrophage proliferation ability of the bioactive peptide SAPRHGSLGFLPRK
[0077] 1. Experimental reagents and instruments
[0078] Reagents: experimental animals balb / c mice (male 6-8 weeks old) Animal Experiment Center, School of Agriculture and Biology, Shanghai Jiaotong University; the mouse spleen lymphocyte-derived bioactive peptide SAPRHGSLGFLPRK obtained in Example 1; 3-(4,5 -Dimethylthiazole-2)-2,5-diphenyltetrazolium bromide (MTT) Amresco; LPS (Lipopolysaccharide) Sigma; Bovine Serum Albumin (BSA) Genebase; Triplex Dissolving solution, an aqueous solution containing 10% SDS, 5% isobutanol and 0.012 mol / L HCl.
[0079] 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 CO 2 Incubator Heraeus Company; Dragon Wellsc...
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