Bioactive polypeptide YFGSGFAAPFFIVRHQLLKK and preparation method and use thereof
A bioactive peptide, bioactive peptide technology, applied in the protein field, to promote lymphocyte proliferation, improve the ability, and reduce the body's morbidity.
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Embodiment 1
[0033] The artificial synthesis of embodiment 1 active peptide YFGSGFAAPFFIVRHQLLKK
[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] 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.
[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.
[0039] ...
Embodiment 2
[0076] The immune activity experiment of embodiment 2 bioactive peptides
[0077] 1. In vitro lymphocyte proliferation ability experiment of biologically active polypeptide YFGSGFAAPFFIVRHQLLKK (MTT method)
[0078] 1. Experimental materials and instruments:
[0079] Reagents and materials: 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 YFGSGFAAPFFIVRHQLLKK obtained in Example 1; mouse lymphocytes Cell extract (purchased from Suo Laibao); RPMI1640 medium (purchased from GIBCO); 3-(4,5-dimethylthiazole-2)-2,5-diphenyltetrazolium bromide ( MTT, purchased from Amresco); concanavalin (ConA, purchased from Sigma); bovine serum albumin (BSA, purchased from Genebase); pepsin (purchased from Sigma); from AB Company).
[0080] Equipment: LRH-250F biochemical incubator, Shanghai Heng Technology Co., Ltd.; GL-22M high-speed refrigerat...
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