Federalnoe gosudarstvennoe byudzhetnoe uchrezhdenie nauki Institut kletochnogo i vnutrikletochnogo simbioza Uralskogo otdeleniya Rossijskoj akademii nauk
发明人:
Sycheva Mariya Viktorovna,Сычева Мария Викторовна,Kartashova Olga Lvovna,Карташова Ольга Львовна,Rogozhin Evgenij Aleksandrovich,Рогожин Евгений Александрович,Peshkova Yuliya Igorevna,Пешкова Юлия Иго
申请号：
RU2016122336
公开号：
RU0002645070C2
申请日:
2016.06.06
申请国别(地区):
RU
年份:
2018
代理人:
摘要：
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.2 tbl, 1 exИзобретение относится к области медицины, а именно к микробиологии и биохимии, и предназначено для получения антимикробных пептидов. Для получения антимикробных пептидов из тромбоцитов курицы домашней осуществляют следующие сбор свежей крови кур в пластиковые контейнеры с добавлением 3,8% раствора цитрата натрия (рН 3,5). Центрифугируют
