% of GFP positive cells 24 hpt

% of GFP positive cells 48 hpt

Time (mpt)

% of GFP positive cells

Time (hpt)

Relative fiuorescence units (R.F.U.)

Relative fiuorescence units 48 hpt

10

15

20

25

5

0

00

5

4500

4000

3500

3000

2500

2000

1500

1000

500

20

15

10

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)

(j)

FIGURE 10.4 (a) Percentage of GFP positive cells depending on the contact time allowed between cells and complexes (b) production of VLPs (in

relative fluorescence units) attained. Percentage of cells attached to a Cy3 complex, expressing GFP or Cy3 positive and expressing GFP protein at 0hpt

(c), 4hpt (e), 10hpt (10hpt), and 48hpt (f). Confocal image showing the cell membrane in red (dyed with cell mask), the nucleus in blue (dyed with

Hoescht) and the expression of the Gag-GFP VLPs (observed in green) in the cytoplasm of the cells at 24hpt (g) and colocalizing at the cell membrane

(observed in yellow) at 48 hpy (h). The colocalization can also be observed by representing the intensity of the different channels (i, j).

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Bioprocessing of Viral Vaccines