Synthetic peptide and application thereof
A technique for synthesizing peptides and wound healing agents, applied in the field of biology, can solve the problems of natural antibacterial peptide content, activity, activity-dependent structure, production cost, etc., achieve significant wound healing effect, realize the effect of low dosage of production and use
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Embodiment 1
[0027] Preparation and purity detection of embodiment 1 synthetic peptide
[0028] In this example, the English parameters involved are well known to those skilled in the field of analytical chemistry, and reference can be made to "Handbook of Analytical Chemistry (6) Liquid Chromatographic Analysis" edited by Zhang Yukui. In addition, TFA refers to trifluoroacetic acid, Acetonitrile refers to acetonitrile, and HCOOH formic acid; the concentration of the mobile phase involved is the volume concentration.
[0029] a chemical synthesis
[0030] The synthetic peptide was chemically synthesized by Shanghai Sangon Bioengineering Technology Service Co., Ltd., purified and confirmed by HPLC.
[0031] Two HPLC analysis of the high-affinity binding polypeptide
[0032] 1 The chromatographic conditions are as follows:
[0033] Measurement: Peak Area;
[0034] Run Time: 30min;
[0035] Calculation Type: Percent;
[0036] Wavelength: 220nm;
[0037] Flow rate: 1.0ml / min;
[0038] ...
Embodiment 2
[0047] The antibacterial activity of embodiment 2 synthetic peptides
[0048] The following strains were purchased from China Institute for the Control of Biological Products.
[0049] The antibacterial activity of antimicrobial peptides was determined according to the following steps:
[0050] LfcinB4-9 was used as a control antimicrobial peptide. Use distilled water to prepare the antimicrobial peptide into a 2560 μg / mL stock solution in a polypropylene centrifuge tube, then dilute it with an equal amount of sterile 0.02% acetic acid (containing 0.4% BSA by volume fraction), and then use a volume fraction of 0.01% acetic acid (containing 0.2% BSA solution by volume fraction) is carrying out a series of double dilutions to the solution after equivalent dilution to obtain a mass concentration of 1280, 640, 320-2.5 μg / mL (successive double dilution ) serial dilutions. Store at 4°C for later use.
[0051] Cultivate the bacteria to be tested on a sterilized broth agar plate o...
Embodiment 3
[0056] Example 3 Application of Synthetic Peptides in Promoting Wound Healing
[0057] 1 Establishment of diabetic mouse model
[0058] Select 8-week-old Kunming rats, fast for 12 hours, and inject 1% streptozotocin solution intraperitoneally at a dose of 120 mg / kg, and fast for 12 hours on the 3rd and 7th days after the injection, respectively. The blood glucose concentration of fasting mice was measured by venous blood collection, and those whose blood glucose concentration exceeded 11.1mmol / L for two consecutive times were regarded as successful establishment of the mouse diabetes model. The mice whose blood glucose concentration reached the requirement were used to establish the trauma model.
[0059] 2 Establishment of mouse trauma model
[0060] Intraperitoneal injection of 7g / L pentobarbital sodium anesthetized mice with 0.7g / kg dose, depilated with 8% sodium sulfide, and wiped with warm water for 2-3 minutes so as to wash off the coat taken off at this position. In ...
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