The Use of Recombinant Pseudotype Virus-Like Particles Harbouring Inserted Target Antigen to Generate Antibodies against Cellular Marker p16INK4A
Figure 2
Analysis of the expression of VP1/VP2-p16INK4A pseudotype VLPs in yeast. (a) Coomassie blue-stained SDS-PAGE. (b) Western blot with the MAb 3D10 against VP1 protein. (c) Western blot with polyclonal antibody against VP2 protein. The same samples were run on each gel. In lanes, there are (1) negative control sample from crude lysate of S. cerevisiae cells transformed with the empty vector pFGG3, (2) crude lysate of yeast transformed with pFGG3-VP1/VP2-p16 plasmid, (3) the soluble fraction recovered after centrifugation of crude lysate of yeast transformed with pFGG3-VP1/VP2-p16 plasmid, (4) VLPs consisting of VP1 protein and fusion protein VP2-p16 purified using sucrose and CsCl gradients, (5) VLPs consisting of VP1 protein and nonmodified VP2 protein purified using sucrose and CsCl gradients, and (6) prestained protein molecular mass marker (Thermo Scientific Fermentas). (d and e) Electron microscopy pictures of VP1/VP2-p16INK4A (d) and nonmodified VP1/VP2 (e) pseudotype VLPs, stained with 2% aqueous uranyl acetate solution and examined by Morgagni 268 electron microscope.
