PMID- 30728922
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20200930
IS  - 2001-3078 (Print)
IS  - 2001-3078 (Electronic)
IS  - 2001-3078 (Linking)
VI  - 8
IP  - 1
DP  - 2019
TI  - An improved 96 well plate format lipid quantification assay for standardisation 
      of experiments with extracellular vesicles.
PG  - 1565263
LID - 10.1080/20013078.2019.1565263 [doi]
LID - 1565263
AB  - The field of extracellular vesicles (EVs) is an exponentially growing segment of 
      biomedical sciences. However, the problems of normalisation and quantification of 
      EV samples have not been completely solved. Currently, EV samples are 
      standardised on the basis of their protein content sometimes combined with 
      determination of the particle number. However, even this combined approach may 
      result in inaccuracy and overestimation of the EV concentration. Lipid bilayers 
      are indispensable components of EVs. Therefore, a lipid-based quantification, in 
      combination with the determination of particle count and/or protein content, 
      appears to be a straightforward and logical approach for the EV field. In this 
      study, we set the goal to improve the previously reported sulfo-phospho-vanillin 
      (SPV) lipid assay. We introduced an aqueous phase liposome standard (DOPC) to 
      replace the purified lipid standards in organic solvents (used commonly in 
      previous studies). Furthermore, we optimised the concentration of the vanillin 
      reagent in the assay. We found that elimination of organic solvents from the 
      reaction mixture could abolish the background colour that interfered with the 
      assay. Comparison of the optimised assay with a commercial lipid kit (based on 
      the original SPV lipid assay) showed an increase of sensitivity by approximately 
      one order of magnitude. Thus, here we report a quick, reliable and sensitive test 
      that may fill an existing gap in EV standardisation. When using the optimised 
      lipid assay reported here, EV lipid measurements can be more reliable than 
      protein-based measurements. Furthermore, this novel assay is almost as sensitive 
      and as easy as measuring proteins with a simple BCA test.
FAU - Visnovitz, Tamas
AU  - Visnovitz T
AD  - Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 
      Hungary.
FAU - Osteikoetxea, Xabier
AU  - Osteikoetxea X
AD  - Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 
      Hungary.
FAU - Sodar, Barbara W
AU  - Sodar BW
AD  - Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 
      Hungary.
FAU - Mihaly, Judith
AU  - Mihaly J
AD  - Institute of Materials and Environmental Chemistry, Research Centre for Natural 
      Sciences of the Hungarian Academy of Sciences, Budapest, Hungary.
FAU - Lorincz, Peter
AU  - Lorincz P
AUID- ORCID: 0000-0001-7374-667X
AD  - Department of Anatomy, Cell and Developmental Biology, Eotvos Lorand University, 
      Budapest, Hungary.
FAU - Vukman, Krisztina V
AU  - Vukman KV
AD  - Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 
      Hungary.
FAU - Toth, Eszter Agnes
AU  - Toth EA
AD  - Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 
      Hungary.
FAU - Koncz, Anna
AU  - Koncz A
AD  - Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 
      Hungary.
FAU - Szekacs, Inna
AU  - Szekacs I
AD  - Nanobiosensorics Laboratory MTA-EK-MFA, Budapest, Hungary.
FAU - Horvath, Robert
AU  - Horvath R
AD  - Nanobiosensorics Laboratory MTA-EK-MFA, Budapest, Hungary.
FAU - Varga, Zoltan
AU  - Varga Z
AD  - Institute of Materials and Environmental Chemistry, Research Centre for Natural 
      Sciences of the Hungarian Academy of Sciences, Budapest, Hungary.
FAU - Buzas, Edit I
AU  - Buzas EI
AUID- ORCID: 0000-0002-3744-206X
AD  - Department of Genetics, Cell- and Immunobiology, Semmelweis University, Budapest, 
      Hungary.
AD  - MTA-SE Immune-Proteogenomics Extracellular Vesicle Research Group, Budapest, 
      Hungary.
LA  - eng
PT  - Journal Article
DEP - 20190129
PL  - United States
TA  - J Extracell Vesicles
JT  - Journal of extracellular vesicles
JID - 101610479
PMC - PMC6352952
OTO - NOTNLM
OT  - EV
OT  - apoptotic bodies
OT  - exosomes
OT  - large oncosomes
OT  - lipid assay
OT  - liposome standard
OT  - microvesicles
OT  - normalisation and quantification
OT  - standardisation
OT  - sulfo-phospho-vanillin (SPV) reaction
EDAT- 2019/02/08 06:00
MHDA- 2019/02/08 06:01
PMCR- 2019/01/29
CRDT- 2019/02/08 06:00
PHST- 2018/07/27 00:00 [received]
PHST- 2018/12/10 00:00 [revised]
PHST- 2019/01/02 00:00 [accepted]
PHST- 2019/02/08 06:00 [entrez]
PHST- 2019/02/08 06:00 [pubmed]
PHST- 2019/02/08 06:01 [medline]
PHST- 2019/01/29 00:00 [pmc-release]
AID - 1565263 [pii]
AID - 10.1080/20013078.2019.1565263 [doi]
PST - epublish
SO  - J Extracell Vesicles. 2019 Jan 29;8(1):1565263. doi: 
      10.1080/20013078.2019.1565263. eCollection 2019.