A kind of bioactive polypeptide fspankkltpkky and its preparation method and application
A technology of biologically active peptides and biologically active peptides, applied in the field of protein, to achieve the effect of promoting the increase of induction amount, improving the ability and improving the quality
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Embodiment 1
[0032] Embodiment 1 The artificial synthesis of active peptide FSPANKKLTPKKY
[0033] 1. Synthesis of Bioactive Peptides
[0034] Synthetic bioactive peptide FSPANKKLTPKKY.
[0035] 2. Confirmation of Bioactive Peptides
[0036] 1) UPLC analysis
[0037] The UPLC conditions are as follows:
[0038] Instruments: Waters ACQUITY UPLC ultra-high performance liquid phase, electrospray, quadrupole, time-of-flight mass spectrometer
[0039] Column specification: BEH C18 column
[0040] Flow rate: 0.4mL / min
[0041] Temperature: 50°C
[0042] UV detection wavelength: 210nm
[0043] Injection volume: 2μL
[0044] Gradient conditions: liquid A: water containing 0.1% formic acid (v / v), liquid B: acetonitrile containing 0.1% formic acid (v / v)
[0045] Time(min) %A %B
0 95.0 5.0
1.50 80.0 20.0
3.50 60.0 40.0
5.00 40.0 60.0
7.00 15.0 85.0
8.00 0.0 100.0
11.00 0.0 100.0
11.50 95.0 5.0
13.00 95.0 5.0
[0046] 2) Mass spectrometry analysis
[0047] The mass spectro...
Embodiment 2
[0061] Example 2 Immune Activity Experiment of Bioactive Peptides
[0062] 1. Determination of the amount of bioactive polypeptide FSPANKKLTPKKY that promotes the induction of nitric oxide in macrophages (Griess method)
[0063] 1. Experimental reagents and instruments:
[0064] Reagents: experimental animals balb / c mice (male 6-8 weeks old) spleen lymphocyte-derived biologically active peptide FSPANKKLTPKKY; LPS, purchased from Sigma; neutral red staining solution, produced by Beyond Biotechnology Research Institute.
[0065] 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; DragonWellscan MK3 microplate reader Labsystems.
[0066] 2. Test method:
[0067] The number of cells added is 2×10 6 / ml cell suspension 100μl / well, after adherent purification, add 200μl / well of RPMI1640 complete culture solution (10% FBS) ...
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