Antibacterial peptide and mitochondrial DNA complex, polyclonal antibody of complex and application of polyclonal antibody
A polyclonal antibody and antimicrobial peptide technology, applied in the field of medicine, can solve the problem of increased risk and achieve good targeting effect, high potency and good antibody quality
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[0025] In the present invention, the complex of the antimicrobial peptide and mitochondrial DNA is preferably a liquid preparation, and the solvent of the liquid preparation is preferably a phosphate buffer; the pH value of the phosphate buffer is preferably 7.4; in the present invention, The concentration of the antimicrobial peptide and mitochondrial DNA complex in the liquid preparation is 1.0-1.5 mg / mL, more preferably 1.2 mg / mL. In the present invention, the preparation method of the liquid preparation preferably includes the following steps: dissolving the complex of the antibacterial peptide and mitochondrial DNA in phosphate buffer, and incubating at 35-38° C. for 20-40 min. In the specific implementation process of the present invention, it is preferred to dissolve the antimicrobial peptide and mitochondrial DNA in phosphate buffer and incubate, the temperature of the incubation is preferably 37°C, and the incubation time is preferably 25-35min, more preferably 30min....
Embodiment 1
[0030] Mitochondrial DNA (mtDNA) Extraction:
[0031] Animal tissue / cell mitochondrial DNA extraction kit (GMS20023.3v.A, GENMED) was used to extract mtDNA from human embryonic kidney cells (HEK293). For details, see the kit instructions. The concentration of the obtained mtDNA was detected by a spectrophotometer (Nano, MestroGen), and the next experiment was carried out after confirming that there was no protein and DNA cross-contamination. (Purity requirements: A260 / A280>1.8, A260 / A280>2.0).
[0032] Peptide synthesis:
[0033] Solid-phase synthesis of peptides (entrusted to biotechnology companies):
[0034] LL-37: LLGDFFRKSKEKIGKEFKRIVQRIKDFLRNLVPRTES
[0035] Peptide Cramp: GLLRKGGEKIGEKLKKIGQKIKNFFQKLVPQPE
[0036] The synthesizer was type 433A (ABI, USA). The correctness of the synthesized sequence was double verified by reversed-phase high-performance liquid chromatography (RP-HPLC) and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (M...
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