Figure 1.

Western blot analysis of LASV antigen positive and negative patient sera and controls for GP1, GP2, and NP proteins. Twenty μL of a 1:4 dilution of precipitated suspected LASV patient sera were resolved per lane of a reducing SDS-PAGE gel, blotted and probed with α-GP1, α-GP2, or α-NP antibodies. In three samples NP, GP1, GP2 were detected, indicating presence of LASV virions (G692-1, G762-1, G765-1; NP, GP1,2 (+)). In G610-3, G676-A, G583-1, and G755-1 only GP1 was detected (GP1 (+)). Only low levels of NP were detected in G787-1 (NP (+)). Whereas in G337-1 and G079-3 only GP1 was detectable (GP1 (+)), GP2 levels were not determined (nd). Similarly, in G090-3 only low levels of NP were detected, but GP2 levels were not determined (nd). A representative suspect LASV patient serum sample that did not reveal detectable levels of all three viral antigens (G543-3) is shown alongside normal uninfected controls (BOM011, BOM019) (NP, GP1,2 (-)). In vitro controls derived from transfection of HEK-293T/17 cells with pcDNA vector (pcDNA) or a LASV GPC construct harvested at 36 hours (GPC 36 h) are shown. Soluble GP1 can be precipitated from the supernatants of cells expressing GPC (GPC 36 h). LASV VLP containing NP, GP1, and GP2 are shown for protein size comparison (L VLP) (in vitro ctrls). Molecular weights for each LASV protein are shown to the left of blots, and identified on the right.