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US 8,263,413 B1Granted
Methods for analyzing lipids and membrane proteins in biological matter using stable isotopes and mass spectrometry
OwnerAndrew S Hanseninventor
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Abstract
Methods for accurately comparing the levels of ionizable lipids in two cell populations that differ in some respect from each other using mass spectroscopy and isotopic labeling are provided. The methods can be used to identify a change in a lipid of interest in response to a cellular, chemical, genetic, or environmental change to the cell population (i.e., a lipid response to a cell perturbation). The change in the lipid of interest can be a change in composition, rate of synthesis, and/or location of the lipid.
Claims12
A method for determining the relative abundance of a lipid of interest, comprising:
- (i) culturing a first sample of a cell population in a first medium, the first medium including a lipid precursor having a first abundance of at least one isotope;
- (ii) culturing a second sample of the cell population in a second medium, the second medium including the lipid precursor with a second and different abundance of the at least one isotope,
- wherein the lipid precursor in each of the samples is incorporated into a biological lipid within the respective samples;
- (iii) modulating one of the samples of the cell population;
- (iv) isolating one or more lipids from each of the first and second samples, the lipids from each of the samples including the biological lipid;
- (v) combining at least a portion of the first sample and a portion of the second sample to form a combined sample, the combined sample comprising the biological lipids from each of the samples;
- (vi) subjecting the isolated lipids in the combined sample to mass-spectrometry;
- and
- (vii) determining the relative abundance of a lipid of interest in the first sample compared to the second sample.
6 more elements
Dependencies are inferred from the claim text.