Author: Lv, Lishan; Li, Xiaoming; Liu, Genmei; Li, Ran; Liu, Qiliang; Shen, Huifang; Wang, Wei; Xue, Chunyi; Cao, Yongchang
Title: Production and immunogenicity of chimeric virus-like particles containing the spike glycoprotein of infectious bronchitis virus Document date: 2014_6_16
ID: k785hl1r_13
Snippet: To produce chimeric VLPs containing the fusion proteins NA/S1 and M1, sf9 cells were co-infected with recombinant baculoviruses expressing NA/S1 and M1 at a MOI of 5. Culture supernatants were harvested at 72 h post-infection. After removing cells by centrifugation at 2,000 × g for 30 min, the VLPs in the supernatants were pelleted by ultracentrifugation at 80,000 × g for 2 h and then resuspended in PBS. The particles were further purified thro.....
Document: To produce chimeric VLPs containing the fusion proteins NA/S1 and M1, sf9 cells were co-infected with recombinant baculoviruses expressing NA/S1 and M1 at a MOI of 5. Culture supernatants were harvested at 72 h post-infection. After removing cells by centrifugation at 2,000 × g for 30 min, the VLPs in the supernatants were pelleted by ultracentrifugation at 80,000 × g for 2 h and then resuspended in PBS. The particles were further purified through a 20-30-40-50% discontinuous sucrose gradient at 80,000 × g for 2 h at 4 o C, after which the purified VLPs were collected and the presence of fusion protein NA/S1 and M1 protein was analyzed by western blot as described above.
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