To Combat Antimicrobial Resistance 20172021 FY of your Ministry of Agriculture, Forestry and Fisheries of Japan. This study was also supported in aspect by the OGAWA Science and Technology Foundation and the Morinaga Foundation for Wellness and Nutrition.PF10.08 PF10.Evaluation from the CD49f/Integrin alpha-6 Proteins Gene ID 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 acquiring extracellular vesicles from cow’s milk Mai Morozumia, Hirohisa Izumib, Muneya Fc-gamma Receptor I/CD64 Proteins Purity & Documentation Tsudac, Takashi Shimizua and Yasuhiro TakedaaaMorinaga Milk Market 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 objective of this study was to evaluate the effects of unique acidification treatments around 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 from the raw milk by ultracentrifugation (UC), therapy with hydrochloric acid, or remedy with acetic acid, followed by filtration and preparation of your whey. The protein concentration with the whey was determined by spectrophotometry, and also the size and concentration of EVs were measured by tunable resistive pulse sensing analysis. Surface marker proteins of EVs were detected by western blot (WB) analysis using the primaryIntroduction: MicroRNAs (miRNAs) are present in lots of foods including milk, which may be involved in various bioactivities when taken orally. Milk consists mostly of two fractions, i.e. casein and whey, and most of the milk miRNAs are thought to be included in extracellular vesicles (EVs) in whey fraction. Biological roles of milk miRNAs will not be fully elucidated and as a result need further investigation. However, procedures for isolating milk-derived EVs (M-EVs) haven’t totally established. The aim of this study was to compare strategies for isolating M-EVs. Approaches: Aiming to reduce the contamination of casein in whey fraction, which is the great obstacle to figuring out M-EVs purity, whey fraction was separated from milk (defatted) by centrifugation only, acetic acid precipitation, or EDTA precipitation (n = 3). M-EVs have been then isolated from each 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). Outcomes: Acetic acid precipitation prevented casein contamination to higher extents. Three combinations, for example “acetic acid precipitation and qEV”, “acetic acid precipitation and EVSeocondL70” and “EDTA precipitation and qEV” had been able to collect bigger numbers of total M-EVs particles than the other combinations. Among the 3 combinations, “EDTA precipitation and qEV” achieved collecting the biggest quantity of M-EVs but “acetic acid precipitation and EVSeocondL70” was capable to obtain M-EVs fractions with high concentration. Summary/Conclusion: The combination of “EDTA precipitation and qEV” is suited to gather the biggest amount of M-EVs. The.