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利用疏水相互作用色谱法从家禽中纯化抗禽流感H5N1的免疫球蛋白蛋黄

Purification immunoglobulin yolk anti avian influenza H5N1 in poultry using hydrophobic interaction chromatography.

作者信息

Prayugo Armanda Dwi, Subroto Toto, Arnafia Wyanda, Widayat Wahyu, Reisinta Dinda, Jauhari Akmal, Munawaroh Siti, Sibit Denniswara, Idar Idar

机构信息

Department of Biomedical Science, Veterinary Medicine Program, Faculty of Medicine, Universitas Padjadjaran, Sumedang, West Java, 45363, Indonesia.

Department of Chemistry, Faculty of Mathematics and Natural Sciences, Universitas Padjadjaran, Sumedang, West Java, 45363, Indonesia; Research Center for Molecular Biotechnology and Bioinformatics, Universitas Padjadjaran, West Java, 40132, Indonesia.

出版信息

Poult Sci. 2025 Feb;104(2):104776. doi: 10.1016/j.psj.2025.104776. Epub 2025 Jan 4.

Abstract

Avian influenza is a significant threat to the poultry industry, and it has become an outbreak in many countries because of its mortality and morbidity. Concerns about the history of avian influenza outbreaks has prompted all countries to enhance their independence in pharmaceutical and biological components as a preparedness measure for any potential occurrences. The production of antibodies such as IgY is a potential alternative. The potential of IgY biomaterials can be utilized as bioreceptors in diagnostic tools. This research aimed to purify IgY anti-avian influenza H5N1 produced from chicken eggs. IgY anti-avian influenza H5N1 can be obtained from chickens vaccinated at 0.5 ml per bird dose using inactivated H5N1 vaccine with a purification stage from graded PEG, hydrophobic interaction chromatography, and concentrated using an ultrafilter. The present study shows that the IgY anti-avian influenza H5N1 can be obtained from chickens vaccinated with an inactivated H5N1 vaccine, from 4 chicken eggs with a total protein content of 40.01 mg, it has biological activity with HI titer activity of 4,915.2 HI units and a recovery value of 24 % with hydrophobic interaction chromatography. Therefore, the purification process with hydrophobic interaction chromatography does not remove the IgY biological antibody. Further development will be required to perform antibody potency as a bioreceptor for diagnostic purposes.

摘要

禽流感对家禽业构成重大威胁,因其致死率和发病率已在许多国家引发疫情。对禽流感疫情历史的担忧促使各国加强在制药和生物成分方面的独立性,作为应对任何潜在疫情的防范措施。生产诸如卵黄免疫球蛋白(IgY)之类的抗体是一种潜在的替代方法。IgY生物材料的潜力可被用作诊断工具中的生物受体。本研究旨在纯化从鸡蛋中产生的抗禽流感H5N1的IgY。抗禽流感H5N1的IgY可从每只鸡接种0.5毫升灭活H5N1疫苗的鸡中获得,并经过分级聚乙二醇(PEG)纯化、疏水相互作用色谱法纯化,然后使用超滤进行浓缩。本研究表明,抗禽流感H5N1的IgY可从接种灭活H5N1疫苗的鸡中获得,从4个鸡蛋中获得的总蛋白含量为40.01毫克,其具有生物活性,血凝抑制(HI)效价活性为4915.2个HI单位,通过疏水相互作用色谱法的回收率为24%。因此,疏水相互作用色谱法的纯化过程不会去除IgY生物抗体。为了将该抗体作为诊断用途的生物受体发挥效力,还需要进一步研发。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4bf1/11761928/1de21304249c/gr1.jpg

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