Fig. 1: Production of LASV GP VLPs for use as immunogens. | npj Vaccines

Fig. 1: Production of LASV GP VLPs for use as immunogens.

From: Adjuvant formulated virus-like particles expressing native-like forms of the Lassa virus envelope surface glycoprotein are immunogenic and induce antibodies with broadly neutralizing activity

Fig. 1

a Schematic representation of LASV GPC. The glycoprotein precursor GPC consists of the stable signal peptide (SSP) (amino acids 1–58), the receptor-binding subunit GP1 (amino acids 59–259), and the fusion-mediating subunit GP2 (amino acids 260–491) containing a transmembrane domain (TM) and a cytoplasmic domain (CD). The signal peptidase (SPase) cleavage site (after amino acid 58), the SKI-1/S1P cleavage site (after amino acid 259), and potential glycosylation sites (Y) are indicated. b Western blot analysis of LASV GP VLPs purified from the cell culture supernatant of MDCK II cells stably expressing LASV GP using primary antibodies against GP1 (mouse AC1) and GP2 (rabbit α4). c Electron micrograph of negatively stained, purified GP VLPs. Scale bar, 100 nm. d Western blot analysis of LASV GP after treatment with endoglycosidases. LASV GP VLPs released from MDCK II LASV GP cells and authentic LASV purified from infected MDCK II cells, HuH7 cells, or BHK cells were treated with either endo-β-N-acetylglucosaminidase H (EndoH) or N-glycosidase F (PNGaseF). GP subunits were detected with primary antibodies against GP1 (mouse AC1) and GP2 (rabbit α4), respectively. e Schematic diagram of the immunization protocol. New Zealand White rabbits were immunized intramuscularly with 300 μg VLPs mixed with Sigma Adjuvant and boosted 28, 49, and 70 days after the first immunization. Serum samples were taken at day 0 and immediately before each boost. Final blood collection was performed on day 77.

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