To Combat Antimicrobial Resistance 20172021 FY from the Ministry of Agriculture, Forestry and Fisheries of Japan. This study was also supported in aspect by the OGAWA Science and Technologies Foundation plus the Morinaga Foundation for Wellness and Nutrition.PF10.08 PF10.Evaluation on the effects of acidification on isolation of extracellular vesicles from bovine milk Md. Matiur Rahmana, Kaori Shimizub, Marika Yamauchic, Ayaka Okadab and Yasuo Inoshimab The United Graduate School of Veterinary Sciences, Gifu University, Gifu, Japan; bGifu University, Gifu, Japan; cGifu University, Gifu, USAaComparison of isolating strategy for getting extracellular vesicles from cow’s milk Mai Morozumia, Hirohisa Izumib, Muneya Tsudac, Takashi Shimizua and Yasuhiro TakedaaaMorinaga Milk Sector Co., Ltd., Zama-City, Japan; bMorinaga Milk Industry Co., Ltd., Zama-city, Japan; cMorinaga Milk Industry Co., Ltd., Zama, JapanIntroduction: Acidification has shown possible for separating casein from raw bovine milk to facilitate isolation and purification of extracellular vesicles (EVs). The goal of this study was to evaluate the effects of distinct acidification treatments on the yield and surface marker proteins of EVs from raw bovine milk. Strategies: Fresh raw bulk milk was collected from healthful dairy cows. Casein was separated in the raw milk by ultracentrifugation (UC), therapy with hydrochloric acid, or treatment with acetic acid, followed by filtration and preparation in the whey. The protein concentration with the whey was determined by spectrophotometry, along with the size and concentration of EVs were measured by tunable resistive pulse sensing analysis. Surface marker proteins of EVs have been detected by western blot (WB) analysis making use of the primaryIntroduction: MicroRNAs (miRNAs) are present in a lot of foods such as milk, which may very well be involved in various bioactivities when taken orally. Milk consists mainly of two fractions, i.e. casein and whey, and most of the milk miRNAs are thought to become incorporated in extracellular vesicles (EVs) in whey fraction. Biological roles of milk miRNAs are usually not totally elucidated and as a result call for further investigation. However, procedures for isolating milk-derived EVs (M-EVs) have not fully established. The aim of this study was to evaluate procedures for isolating M-EVs. Procedures: Aiming to reduce the BTLA Proteins Biological Activity contamination of casein in whey fraction, which is the great obstacle to determining M-EVs CD49c/Integrin alpha-3 Proteins Recombinant Proteins purity, whey fraction was separated from milk (defatted) by centrifugation only, acetic acid precipitation, or EDTA precipitation (n = 3). M-EVs were then isolated from every whey fraction by ultracentrifugation, an exoEasy Maxi kitISEV2019 ABSTRACT BOOK(Qiagen), a qEV kit (Izon Science) or an EVSecondL70 kit (GL Sciences). The number of M-EVs particles was measured applying NanoSight (Malvern Instruments). Results: Acetic acid precipitation prevented casein contamination to higher extents. 3 combinations, like “acetic acid precipitation and qEV”, “acetic acid precipitation and EVSeocondL70” and “EDTA precipitation and qEV” had been in a position to collect bigger numbers of total M-EVs particles than the other combinations. Among the 3 combinations, “EDTA precipitation and qEV” accomplished collecting the largest quantity of M-EVs but “acetic acid precipitation and EVSeocondL70” was able to obtain M-EVs fractions with high concentration. Summary/Conclusion: The combination of “EDTA precipitation and qEV” is suited to gather the largest quantity of M-EVs. The.