APR January/February 2022 - 32

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DRUG DEVELOPMENT
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Figure 4. Autologous Compatible (other than the patient) Cells Donor Therapy
Table 1.
Virus
Adeno-associated Viruses (AAV)
Adenovirus
Alphavirus
Baculovirus
Herpesvirus
Lentivirus
Poxvirus
Reovirus
Retrovirus
Co-transfection of plasmid DNA
Co-transfection of plasmid DNA
In vitro transcription of rec & helper RNA
Co-transfection of plasmid DNA
Virus Stock Preparation
Virus production in HEK293 cells
Virus production in HEK, PerC6 cells
Virus production in insect cells
Co-transfection of amplicon & helper DNA Virus production in Vero cells
Co-transfection of plasmid DNA
Co-transfection of plasmid DNA
Infection with reovirus
Co-transfection of plasmid DNA
Simian Vacuolating Virus 40 (SV40) Co-transfection of plasmid DNA
Harvest of virus stock
Harvest of virus stock
Electroporation of RNA into BHK cells Harvest of virus stock
Harvest of virus stock
Harvest of virus stock
Virus production in HEK293 cells
Virus production in cell lines
Virus production in L929 cells
Virus production in packaging cells
Virus production in COS cells
Harvest of virus stock
Harvest of virus stock
Harvest of virus stock
Harvest of virus stock
Harvest of virus stock
Purification, concentration
Purification, concentration
Purification, only for GMP
Purification, concentration
Total Time
5-6 days
10 days
2-3 days
14 days
Purification, ultracentrifugation 7 days
Purification, concentration
Purification, concentration
Purification, concentration
Purification, concentration
7 days
7 days
3-4 days
10 days
Purification, ultracentrifugation 7 days
Schematic presentation of viral vectors and the time required. The various steps involved in preparation of virus stocks are briefly described. The time required for the preparation
does not include subcloning into vectors and preparation of plasmid DNA. However, these procedures are generally independent of which viral vector system is used.3
and viral delivery. These techniques involve the simultaneous
delivery of two distinct nucleic acids into the same cell and are
often used to achieve stable transfections. Transfections methods
have evolved to include several new methods such as the biolistic
delivery systems that use high velocity microparticles to deliver
nucleic acids into cells, and in vivo transfection protocols that
facilitate systemic delivery of siRNA molecules.
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Viral Transfection
This method involves the use of viral vectors to deliver nucleic acids
into cells. Viral delivery systems such as lentiviral, adenoviral and
oncoretroviral vectors can be used for transferring nucleic acids, even
in hard-to-transfect cells.

APR January/February 2022

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