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. 2016 Aug 12;11(8):e0160995. doi: 10.1371/journal.pone.0160995

Fig 3. Purification profile of V3 infiltrated tobacco plants.

Fig 3

(A) Immunoblot showing the target L1 protein collected from CsCl density gradients top (TB) and lower bands (LB) from V3 infiltrated plants. (B) The size exclusion chromatogram of CsCl collected and dialyzed V3 sample on 16/600 superdex-200 prep grade column using FPLC system. The molecular standards represented on the chromatogram are BD; bluedex (2000 kDa), C; conalbumin (75 kDa), O; ovalbumin (43 kDa). (C) SDS-PAGE and immunoblot showing the L1 band of dissociated chromatographic purified VLPs. The VLPs from insect cells were used as loading positive control and arrows in immunoblots show HPV L1 band at 56 kDa; 16E was used as HPV16 L1 sequential epitopes detecting monoclonal antibody. (D) ELISA profile for characterization of plant purified HPV16 VLPs using monoclonal (16A, C, and E) and polyclonal (R intact) in-house antibodies. 16A and C detect conformational epitopes, whereas 16E and R intact detect both conformational, as well as sequential epitopes. (E) Validation of purified VLPs by immunoblot using the same sequential and conformational antibodies as described in D. One way analysis of variance (ANOVA) was done to appreciate the dose response and a value *p < 0.05 was considered significantly different.