A Single In-Vial Dual Extraction Strategy for the Simultaneous Lipidomics and Proteomics Analysis of HDL and LDL Fractions.
A single in-vial dual extraction (IVDE) procedure for the subsequent analysis of lipids and proteins in the high-density lipoprotein (HDL) and low-density lipoprotein (LDL) fractions derived from the same biological sample is presented. On the basis of methyl-tert-butyl ether (MTBE) extraction, IVDE leads to the formation of three phases: a protein pellet at the bottom, an aqueous phase with polar compounds, and an ether phase with lipophilic compounds. After sample extraction, performed within a high-performance liquid chromatography vial insert, the ether phase was directly injected for lipid fingerprinting, while the protein pellet, after evaporation of the remaining sample, was used for proteomics analysis. Human HDL and LDL isolates were used to test the suitability of the IVDE methodology for lipid and protein analysis from a single sample in terms of data quality and matching composition to that of HDL and LDL. Subsequently, HDL and LDL fractions isolated from ApoE-KO and wild-type mice were used to validate the capacity of IVDE for revealing changes in lipid and protein abundance. Results indicate that IVDE can be successfully used for the subsequent analysis of lipids and proteins with the advantages of time saving, simplicity, and reduced sample amount.
CEMBIO, Centre for Metabolomics and Bioanalysis, San Pablo CEU University , Madrid 28003, Spain.
Joanna
J
Godzien
Clinical Research Centre, Medical University of Bialystok , Bialystok 12-089, Poland.
Michal
M
Ciborowski
CEMBIO, Centre for Metabolomics and Bioanalysis, San Pablo CEU University , Madrid 28003, Spain.
Emily Grace
EG
Armitage
Cardiovascular Proteomics Laboratory, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC) , 28029 Madrid, Spain.
Inmaculada
I
Jorge
Cardiovascular Proteomics Laboratory, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC) , 28029 Madrid, Spain.
Emilio
E
Camafeita
Vascular Research Laboratory, IIS-Fundación Jiménez Díaz-Autonoma University , 28040 Madrid, Spain.
Elena
E
Burillo
Vascular Research Laboratory, IIS-Fundación Jiménez Díaz-Autonoma University , 28040 Madrid, Spain.
Jose Luis
JL
Martín-Ventura
CEMBIO, Centre for Metabolomics and Bioanalysis, San Pablo CEU University , Madrid 28003, Spain.
Francisco J
FJ
Rupérez
Cardiovascular Proteomics Laboratory, Centro Nacional de Investigaciones Cardiovasculares Carlos III (CNIC) , 28029 Madrid, Spain.
Jesús
J
Vázquez
CEMBIO, Centre for Metabolomics and Bioanalysis, San Pablo CEU University , Madrid 28003, Spain.
Coral
C
Barbas
D000818:Animals; D001057:Apolipoproteins E / Q000235:genetics; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D008055:Lipids / Q000032:analysis*; D008074:Lipoproteins / Q000032:analysis*; D008075:Lipoproteins, HDL / Q000032:analysis; D008077:Lipoproteins, LDL / Q000032:analysis; D008738:Methyl Ethers; D051379:Mice; D018345:Mice, Knockout; D040901:Proteomics / Q000379:methods*; D052616:Solid Phase Extraction
D001057:Apolipoproteins E; D008055:Lipids; D008074:Lipoproteins; D008075:Lipoproteins, HDL; D008077:Lipoproteins, LDL; D008738:Methyl Ethers; C043243:methyl tert-butyl ether
Apo-KO mouse; HDL; LDL; in-vial dual extraction (IVDE); lipid annotation; lipidomics; metabolomics; multiomics; proteomics; single sample extraction