2022
DOI: 10.3390/ijms232416106
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Analysis of Proteins and Peptides of Highly Purified CD9+ and CD63+ Horse Milk Exosomes Isolated by Affinity Chromatography

Abstract: Exosomes are nanovesicles with a 40–150 nm diameter and are essential for communication between cells. Literature data suggest that exosomes obtained from different sources (cell cultures, blood plasma, urea, saliva, tears, spinal fluid, milk) using a series of centrifugations and ultracentrifugations contain hundreds and thousands of different protein and nucleic acid molecules. However, most of these proteins are not an intrinsic part of exosomes; instead, they co-isolate with exosomes. Using consecutive ult… Show more

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Cited by 10 publications
(24 citation statements)
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“…Affinity chromatography ( Burkova et al, 2018 ; Sedykh et al, 2022 ) is a molecular recognition-based separation method that involves affinity chromatography and immunoaffinity chromatography, among others. These techniques use the binding of specific compounds or antibodies to exosome molecules to isolate exosomes, enabling efficient separation and purification.…”
Section: Isolation and Purification Of Lactobacilli Exosomes: Methods...mentioning
confidence: 99%
“…Affinity chromatography ( Burkova et al, 2018 ; Sedykh et al, 2022 ) is a molecular recognition-based separation method that involves affinity chromatography and immunoaffinity chromatography, among others. These techniques use the binding of specific compounds or antibodies to exosome molecules to isolate exosomes, enabling efficient separation and purification.…”
Section: Isolation and Purification Of Lactobacilli Exosomes: Methods...mentioning
confidence: 99%
“…In [78], biocompatible 1% sodium citrate was able to disintegrate casein micelles (70-200 nm in sizes) into 40 nm monomers, allowing for efficient purification of high quantities of both non-exosome and exosome EV subsets by subsequent differential ultracentrifugation. In other study [79], ultracentrifugation was first applied followed by gel filtration that allowed to isolate purified exosomes from co-precipitating proteins and their complexes. The application of affinity chromatography, using Sepharose resin with antibodies against exosome surface protein markers CD9 and CD63, resulted in additional exosome purification [79].…”
Section: Isolation/purification Of Evs From Milkmentioning
confidence: 99%
“…In other study [79], ultracentrifugation was first applied followed by gel filtration that allowed to isolate purified exosomes from co-precipitating proteins and their complexes. The application of affinity chromatography, using Sepharose resin with antibodies against exosome surface protein markers CD9 and CD63, resulted in additional exosome purification [79]. Recently, two protocols have been reported [80] that used the incorporation of solubilization steps, optimized timing, temperature, and divalent cation chelation into either centrifugation workflow or tangential flow filtration workflow.…”
Section: Isolation/purification Of Evs From Milkmentioning
confidence: 99%
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“…The specific morphology of “non-vesicle” lets us classify them as intermediate and low-density lipoproteins (20–40 nm) as well as very low density (40–100 nm). TEM examination of exosome preparations of the female placenta [ 15 , 28 , 29 ] and mare’s milk [ 30 , 31 ], isolated by centrifugation and ultracentrifugation, revealed all types of impurities described in [ 14 ]. Another proof of the need for additional purification of exosome preparations after ultracentrifugation is the detection of up to several thousand different proteins in these preparations [ 11 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 ].…”
Section: Introductionmentioning
confidence: 99%