Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations

High-density lipoprotein (HDL) and exosomes are promising sources of biomarkers. However, the limited sample volume and access to the ultracentrifuge equipment are still an issue during HDL and exosome isolation. This study aimed to isolate HDL and exosomes using an ultracentrifugation-free method w...

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Autores principales:	Rian Ka Praja, Wisitsak Phoksawat, Patcharaporn Tippayawat, Amonrat Jumnainsong, Chanvit Leelayuwat
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	biomarker exosome HDL isolation small serum volume Physics QC1-999 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/3f8ce314bc7040019279ad29b49969aa
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spelling	oai:doaj.org-article:3f8ce314bc7040019279ad29b49969aa2021-11-25T18:59:20ZAlternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations10.3390/separations81102042297-8739https://doaj.org/article/3f8ce314bc7040019279ad29b49969aa2021-11-01T00:00:00Zhttps://www.mdpi.com/2297-8739/8/11/204https://doaj.org/toc/2297-8739High-density lipoprotein (HDL) and exosomes are promising sources of biomarkers. However, the limited sample volume and access to the ultracentrifuge equipment are still an issue during HDL and exosome isolation. This study aimed to isolate HDL and exosomes using an ultracentrifugation-free method with various small serum volumes. HDL was isolated from 200 µL (HDL200) and 500 µL (HDL500) of sera. Three different volumes: 50 µL (Exo50), 100 µL (Exo100), and 250 µL (Exo250) were used for exosome isolation. HDL and exosomes were isolated using commercial kits with the modified method and characterized by multiple approaches. The HDL levels of HDL200 and HDL500 were not significantly different (<i>p</i> > 0.05), with percent recoveries of >90%. HDL200 and HDL500 had the same protein pattern with a biochemical similarity of 99.60 ± 0.10%. The particle sizes of Exo50, Exo100, and Exo250 were in the expected range. All isolated exosomes exhibited a similar protein pattern with a biochemical similarity of >99%. In conclusion, two different serum volumes (200 and 500 µL) and three different serum volumes (50, 100, and 250 µL) can be employed for HDL and exosome isolation, respectively. The possibility of HDL and exosome isolation with small volumes will accelerate biomarker discoveries with various molecular diagnostic approaches.Rian Ka PrajaWisitsak PhoksawatPatcharaporn TippayawatAmonrat JumnainsongChanvit LeelayuwatMDPI AGarticlebiomarkerexosomeHDLisolationsmall serum volumePhysicsQC1-999ChemistryQD1-999ENSeparations, Vol 8, Iss 204, p 204 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	biomarker exosome HDL isolation small serum volume Physics QC1-999 Chemistry QD1-999
spellingShingle	biomarker exosome HDL isolation small serum volume Physics QC1-999 Chemistry QD1-999 Rian Ka Praja Wisitsak Phoksawat Patcharaporn Tippayawat Amonrat Jumnainsong Chanvit Leelayuwat Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations
description	High-density lipoprotein (HDL) and exosomes are promising sources of biomarkers. However, the limited sample volume and access to the ultracentrifuge equipment are still an issue during HDL and exosome isolation. This study aimed to isolate HDL and exosomes using an ultracentrifugation-free method with various small serum volumes. HDL was isolated from 200 µL (HDL200) and 500 µL (HDL500) of sera. Three different volumes: 50 µL (Exo50), 100 µL (Exo100), and 250 µL (Exo250) were used for exosome isolation. HDL and exosomes were isolated using commercial kits with the modified method and characterized by multiple approaches. The HDL levels of HDL200 and HDL500 were not significantly different (<i>p</i> > 0.05), with percent recoveries of >90%. HDL200 and HDL500 had the same protein pattern with a biochemical similarity of 99.60 ± 0.10%. The particle sizes of Exo50, Exo100, and Exo250 were in the expected range. All isolated exosomes exhibited a similar protein pattern with a biochemical similarity of >99%. In conclusion, two different serum volumes (200 and 500 µL) and three different serum volumes (50, 100, and 250 µL) can be employed for HDL and exosome isolation, respectively. The possibility of HDL and exosome isolation with small volumes will accelerate biomarker discoveries with various molecular diagnostic approaches.
format	article
author	Rian Ka Praja Wisitsak Phoksawat Patcharaporn Tippayawat Amonrat Jumnainsong Chanvit Leelayuwat
author_facet	Rian Ka Praja Wisitsak Phoksawat Patcharaporn Tippayawat Amonrat Jumnainsong Chanvit Leelayuwat
author_sort	Rian Ka Praja
title	Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations
title_short	Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations
title_full	Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations
title_fullStr	Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations
title_full_unstemmed	Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations
title_sort	alternative method for hdl and exosome isolation with small serum volumes and their characterizations
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/3f8ce314bc7040019279ad29b49969aa
work_keys_str_mv	AT riankapraja alternativemethodforhdlandexosomeisolationwithsmallserumvolumesandtheircharacterizations AT wisitsakphoksawat alternativemethodforhdlandexosomeisolationwithsmallserumvolumesandtheircharacterizations AT patcharaporntippayawat alternativemethodforhdlandexosomeisolationwithsmallserumvolumesandtheircharacterizations AT amonratjumnainsong alternativemethodforhdlandexosomeisolationwithsmallserumvolumesandtheircharacterizations AT chanvitleelayuwat alternativemethodforhdlandexosomeisolationwithsmallserumvolumesandtheircharacterizations
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Alternative Method for HDL and Exosome Isolation with Small Serum Volumes and Their Characterizations

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