A kind of biologically active peptide rrecpsdecgagvf and its preparation method and application
A bioactive peptide and sequence technology, applied in the protein field, can solve the problem of lack of research, achieve the effect of good anti-inflammatory activity, promote the increase of induction and improve the quality of life
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Embodiment 1
[0032] Example 1 Artificial synthesis of bioactive peptide RRECPSDECGAGVF
[0033] 1. Synthesis of Bioactive Peptides
[0034] Synthetic bioactive peptide RRECPSDECGAGVF.
[0035] 2. Confirmation of Bioactive Peptides
[0036] 1) UPLC analysis
[0037] The UPLC conditions are as follows:
[0038] Instrument: Waters ACQUITY UPLC ultra-high performance liquid phase, electrospray, quadrupole, time-of-flight mass spectrometer
[0039] Column Specifications: BEH C18 Column
[0040] Flow rate: 0.4mL / min
[0041] Temperature: 50℃
[0042] UV detection wavelength: 210nm
[0043] Injection volume: 2μL
[0044] Gradient conditions: solution A: water containing 0.1% formic acid (v / v), solution 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
[0047] The mass spectrometry co...
Embodiment 2
[0061] Example 2 Immunoactivity test of biologically active peptides
[0062] 1. Determination of macrophage nitric oxide induction by bioactive peptide RRECPSDECGAGVF (Griess method)
[0063] 1. Experimental reagents and instruments:
[0064] Reagents: experimental animals balb / c mice (male 6-8 weeks old) spleen lymphocyte-derived bioactive peptide RRECPSDECGAGVF; LPS, purchased from Sigma; neutral red staining solution, produced by Biyuntian Institute of Biotechnology.
[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 6 100 μl / well of cell suspension / ml, and 200 μl / well of peptide-containing RPMI1640 complete culture medium (10% FBS) was added after adherence purification. In the inflam...
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