Figure 1

“Spin-thru” purification isolates mature cores from HIV-1 virions. The cores were isolated from the HIV-1 virions concentrated from culture media of infected Sup-T1, PMA-activated and non-activated THP1 cells by the “spin-thru” purification. A – CA p24^Gag profiles of 30-70% sucrose gradients after centrifugation of concentrated HIV-1 virions (upper panel) and “spin-thru”purified viral cores (lower panel). The 0.4 ml sucrose gradient fractions were collected, dialyzed against PBS and subjected to p24 ELISA. B – Electron microscopy of uranyl acetate negatively stained HIV-1 virions concentrated through 30% sucrose cushion (B1-B3), ultrathin sections of virions harvested from infected Sup-T1 cells (B4) and negatively stained core preparations after “spin-thru” purification (B5-B8). The negatively stained viral particles are indicated by single white arrows; extracellular vesicle contaminants in the preparations of concentrated virions are indicated by double white arrows; mature virions in the sections of viral preparatins are indicated by single black arrows, immature particles – by double black arrows. C. The cores of viruses produced by Sup-T1 and activated THP1 cells do not have differences in the profile of the GagPol processing products. The “spin-thru” purified and p24^Gag normalized cores were analyzed by Western blotting using the human HIV immunoglobulin (HIV-IG) from NIH AIDS Research & Reference Reagent Program. The bands of HIV-1 Gag and Pol processing products are indicated on the left.