Fractionating human plasma into density groups (e.g., HDL) allows for more effective identification of differences in plasma proteins between non-AD patients and AD patients. The current standard for fractionating plasma lipoproteins is ultracentrifugation, however, this is both time and labor intensive and impractical for clinical use. Apolipoprotein AI (ApoAI) is a major protein on HDL. The objective of this study was to develop an immunoprecipitation protocol to isolate HDL from human plasma. ApoAI specific antibodies coupled to magnetic beads were used to precipitate ApoAI from human plasma samples. ApoAI was then eluted from the beads, digested using a trypsin digest protocol, and measured by a liquid chromatography-mass spectrometer (LC-MS). Results from the LC-MS were used to determine the successful isolation of HDL and identify other co-isolated proteins from the plasma. In the future, antibodies specific to other plasma fractions (e.g., chylomicrons, VLDL, IDL, LDL) will be used to isolate those fractions from plasma samples.
This research was supported by the Undergraduate Research Opportunities Program (UROP).
Bhalerao, Aniket; Flaten, Zach; Li, Danni.
Development of Immunoprecipitation Protocols to Isolate Plasma Lipoproteins for use in Characterizing the Alzheimer’s disease Plasma Lipoproteome.
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