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pVAC - Induction of Neutralizing Antibodies
pVAC is a DNA vaccine plasmid family specifically designed to stimulate a humoral immune response by intramuscular injection. Antigenic proteins
are targeted and anchored to the cell surface by cloning the gene of interest in frame between the IL2 signal sequence and the C-terminal
transmembrane anchoring domain of human placental alkaline phosphatase. The antigenic peptide produced on the surface of muscle cells is thought
to be taken up by antigen presenting cells (APCs) and processed through the major histocompatibility complex (MHC) class II pathway1, 2, 3 .
Description
Cell Surface Expression of the Antigenic Protein
Two pVAC backbones are available:
• pVAC1-mcs is designed for the cloning of an antigenic gene that is not
naturally secreted. The MCS is flanked by the 5’ IL2 signal sequence and
the 3’ glycosylphosphatidylinositol (GPI) anchoring domain of placental
alkaline phosphatase (PLAP).
• pVAC2-mcs is designed for the cloning of an antigenic gene that already
possesses a signal sequence. The MCS is located upstream of the GPI
anchoring domain of PLAP.
High-level Expression of the Antigenic Gene
pVAC plasmids utilize the promoter region of the rhesus monkey EF1a
gene to achieve high levels of expression in skeletal muscle cells and antigen
presenting cells. Expression levels are further increased by the addition of
the SV40 enhancer that heightens the ability of the plasmid to be
transported into the nucleus, especially in non-dividing cells.
Sustained Expression of the Antigenic Gene
The bacterial region required for replication and selection of pVAC in
E. coli contains a reduced number of immunogenic CpG motifs by featuring
a minimized origin of replication (Ori) and a CpG-free Zeocin -resistance
gene (Sh-DCpG).
Convenient Cloning of the Antigenic Gene
pVAC contains a multiple cloning site (MCS) with several commonly used
restriction sites.
1. Corr M. et al. 1999. In vivo priming by DNA injection occurs predominantly by antigen
transfer. J Immunol. 163(9):4721-7. 2. Forns X. et al. 1999. DNA immunization of mice and
macaques with plasmids encoding hepatitis C virus envelope E2 protein expressed
intracellularly and on the cell surface. Vaccine 17:1992-2002. 3. McCluskie MJ et al. 1999.
Route and method of delivery of DNA vaccine influence immune responses in mice and
non-human primates. Mol Med 5:287-300.
Contents and Storage
Each pVAC plasmid is provided as 20 μg of lyophilized DNA. Product is
shipped at room temperature and should be stored at -20°C. Plasmid is
stable up to one year when properly stored.
Each pVAC is provided with 4 pouches of E. coli Fast-Media ® Zeo (2 TB
and 2 Agar, see pages 44-45).
Ori
Ori
www.invivogen.com/dna-vaccination
EM7 Sh∆CpG
EF1 pAn
EM7 Sh∆CpG
SV40
SV40
IL2 ss
rh EF1α
Intron
5’- Bam HI, Eco RV, Bgl II, Eco RI -3’
rh EF1α
5’- Bam HI, Eco RV, Bgl II, Eco RI -3’
PRODUCT QTY CAT. CODE
pVAC1-mcs 20 μg pvac1
pVAC2-mcs 20 μg pvac2
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ANTIBODIES & VACCINATION 5