Reptile antibacterial peptide OH-CATH30 nanometer granules and nanofibers and application of nanofibers
A technology of OH-CATH30 and reptiles, which is applied in the field of biomedicine, can solve problems such as medical problems where no treatment has been found, and achieve significant anti-infection effects, good inhibitory effects, and significant antibacterial effects
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Embodiment 1
[0032] Example 1, Preparation of Reptile Cathelicidin Antimicrobial Peptide Nanoparticles and Detection of Encapsulation Efficiency and Slow Release
[0033] 1. Preparation of Reptile Cathelicidin Antimicrobial Peptide Nanoparticles
[0034] The preparation of reptile cathelicidin antimicrobial peptide nanoparticles comprises the following steps:
[0035] (1) Prepare a carboxymethyl chitosan (CMCS) solution with a concentration of 1 mg / mL and an antimicrobial peptide OH-CATH30 solution with a concentration of 1 mg / mL with deionized water, respectively, and filter them with a 0.22um microporous membrane until they are well bacteria in the EP tube.
[0036] The antimicrobial peptide OH-CATH30 solution used in the present invention is referred to as OH-30 solution for short, and the sequence of the antimicrobial peptide OH-30 is: KFFKKLKNSVKKRAKKFFKKPRVIGVSIPF (SEQ ID No: 1).
[0037] (2) Transfer the above-mentioned certain amount of CMCS solution to a sterilized container wit...
Embodiment 2
[0044] Embodiment 2, the preparation of reptile cathelicidin antimicrobial peptide nanofiber
[0045] The preparation method of reptile cathelicidin antimicrobial peptide nanofiber is as follows:
[0046] (1) Dissolve 2% (w / v) chitosan (CS) (0.1 g) in 1 mL of 15% (v / v) acetic acid solution and stir until dissolved.
[0047] (2) 9% (w / v) polyvinyl alcohol (PVA) (1.8 g) was dissolved in 8.4 mL of deionized water and stirred at constant temperature at 95 °C for 1.5 h.
[0048] (3) Add 1.4mL DMSO, 0.1mL glycerin and 0.1mL Tween 80 to the above PVA solution, continue to stir at 80°C for 1 hour and then cool to room temperature.
[0049] (4) Add the Cathelicidin antimicrobial peptide nanoparticles prepared in Example 1 (nanoparticle concentration is 0.1-0.3% (w / v)) into the CS solution, mix well, add to the PVA solution and stir magnetically. After continuing the magnetic stirring for 1 hour, the mixture was left standing to remove air bubbles.
[0050] (5) Finally, the above sol...
Embodiment 3
[0052] Embodiment 3, the effect of reptile cathelicidin antimicrobial peptide OH-CATH30 nanoparticles on different bacteria
[0053] Cultivate the strain to be tested overnight in LB medium at 37°C, inoculate it into fresh LB medium the next day, and continue culturing at 37°C until the logarithmic growth phase (OD 600 about 0.9-1), and then diluted with fresh LB medium to a concentration of about 10 6 cfu / ml, spare. Add 100ul of the diluted bacterial solution obtained above to a new 96-well plate, and then add the pre-prepared antimicrobial peptide OH-CATH30 nanoparticles (the amount of nanoparticles added is pre-calculated according to the encapsulation efficiency , so that the final concentration is 100g / ml) was mixed evenly with the bacterial solution, and a blank control group was set. Incubate at 37°C for 12-14 hours, detect OD with a microplate reader 600 , to calculate the inhibition rate. Each experiment was repeated three times, and the results are shown in ima...
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