Chemical modifications, viral particles

In Section 2, factors that could lead to particle assembly and secretion into the supernatant were discussed. At this point a deeper analysis of the factors affecting cell infection will be made. Optimisation of the production process should take into account virus-cell interactions, and more specifically viral attachment and internalisation into the cell. The impact of chemical modifications of the medium in baculovirus attachment-internalisation has not been carefully studied. It is widely known for example, that serum increases the infec-tivity of baculovirus. These reviewers have had one case where we were only able to succeed in infecting Sf9 cells adapted to growth in serum-free media [52], with a baculovirus produced by Sf9 cells (not adapted to grow in serum-free media), after adding serum to the culture (authors unpublished observations). However, since serum is not desirable for use in industrial production, its utilisation should be avoided as much as possible. 

In detail, viral wild-type particles of one set were labeled with the fluorescent dye AlexaFluor (AF) 488 and biotin both groups were attached to surface available lysines. Another batch was labeled with AF568 and biotin, also at addressable lysines. Both types of building block, in the following referred to as CPMV-biotin-AF488 and CPMV-biotin-AF568, were characterized by TEM, UV-visible spectroscopy, native gel electrophoresis, and dot blot studies. TEM studies verified that the particles remained intact after chemical modification. UV-visible spectroscopy confirmed covalent modification and also allowed quantification of the number of labels per particle the particles displayed around 40 biotin moieties and around 200 dyes. 

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