Biomedical Chemistry: Research and Methods 2026, 9(2), e00_327

PROTOCOL FOR THE PREPARATIVE ISOLATION OF LDL AND HDL FRACTIONS FROM HUMAN BLOOD PLASMA

L.A. Kaluzhskiy, E.O. Yablokov, I.Yu. Toropygin, M.A. Konstantinov, O.V. Gnedenko*, Y.V. Mezentsev, A.S. Ivanov, A.I. Archakov

Institute of Biomedical Chemistry, 10/8 Pogodinskaya str., Moscow, 119121 Russia; *e-mail: gnedenko.oksana@gmail.com

Keywords:blood plasma lipoproteins; LONP, LDL, HDL; high-performance gel chromatography; mass spectrometry identification of apoproteins

DOI:10.18097/BMCRM00327

The whole version of this paper is available in Russian.

The pathogenesis of atherosclerosis, the main cause of cardiovascular disease, including coronary heart disease and stroke, involves plasma lipoproteins, which differ in protein and lipid composition, size, charge and function. At present, chromatographic methods, in particular gel filtration (SEC), are widely used to isolate lipoproteins. In this article, we present a protocol for the preparative isolation of low-density lipoprotein (LDL) and high-density lipoprotein (HDL) fractions from human blood plasma using gel chromatography. To determine the boundaries of the elution zones of apolipoproteins A-1 and B-100, HDL and LDL markers, respectively, we used liquid chromatography with tandem mass spectrometry (LC-MS/MS) with ELISA cross-validation. Lipoprotein fractions obtained with the help of the SEC can be used for various studies of scientific and applied importance.
Figure 1. Chromatogram of human blood plasma and distribution of A-1 and B-100 in fractions. Blue – chromatogram of blood plasma according to optical density data at a wavelength of 280 nm, yellow – distribution of A-1 in chromatographic fractions according to the emPAI value (A) and ELISA (C), pink – distribution of B-100 in chromatographic fractions according to the emPAI value (B) and ELISA (D).

FUNDING

The work was carried out within the framework of the Program for Basic Research in the Russian Federation for a long-term period (2021–2030) (No. 122030100168-2).

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