METHOD FOR ANTIMICROBIAL PEPTIDES PRODUCTION FROM DOMESTIC CHICKEN PLATELETS Russian patent published in 2018 - IPC A61K38/02 A61K35/14 C12P21/00 

FIELD: medicine.

SUBSTANCE: to obtain antimicrobial peptides from the domestic chicken platelets, the following fresh blood of chicken is collected into plastic containers with addition of 3.8% sodium citrate solution (pH 3.5). The collected blood is centrifuged at 250 g for 30 minutes. The resulting supernatant is centrifuged at 1000 g for 30 minutes. The resulting precipitated platelets are washed three times with medium 199 with addition of 3.8% sodium citrate. The washed platelet mass is resuspended in 10% acetic acid and kept at -15…-20 °C for 24 hours. The product is thawed and centrifuged at 1000 g for 40 minutes. Antimicrobial substances are precipitated from the resulting supernatant with chilled acetone in a ratio of 1:7 (v/v) for 12 hours at 4 °C. The precipitate is centrifuged at 6000 g for 10 minutes, dried in air with vigorous stirring and crushed. The resulting acetone precipitate is desalted by reversed-phase high-performance liquid chromatography (RP-HPLC) in a stepwise gradient increasing the concentration of 80% aqueous acetonitrile in 0.1% aqueous trifluoracetic acid (TFA) to a value of 75%. After elution from the column, the total extract is removed from the excess content of acetonitrile on a vacuum centrifuge, followed by lyophilization. The dried desalted extract is diluted again in 0.1% TFA and separated by analytical RP-HPLC in a linear gradient of increasing the concentration of aqueous acetonitrile in aqueous TFA from 8 to 40% (basic) and 40-56% (optional). Antimicrobial peptides are collected and lyophilized.

EFFECT: obtaining of purified antimicrobial peptides from the domestic chicken platelets.

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