2
The viral system includes vectors developed from retrovirus (RV), adenovirus (AdV), adeno-
associated virus (AAV), lentivirus (LV), and herpes simplex virus (HSV). AAV belongs to the family
of Parvoviridae, a group of viruses among the smallest of single-stranded and non-enveloped DNA
viruses. There are eight different AAV serotypes reported to date.
Introduction
Recombinant AAV-2 is the most common serotype used in gene delivery, and can be produced at high
titers with a helper virus or Cell Biolabs’ AAV Helper-Free System. AAV-2 can infect both dividing
and non-dividing cells and can be maintained in the human host cell, creating the potential for long-
term gene transfer. Because AAV-2 is a naturally defective virus, requiring provision of several
factors in
trans
for productive infection, it is considered the safest viral vector to use. Recently a new
vector, AAV-DJ, was developed using DNA family shuffling to create a hybrid capsid from 8 different
AAV serotypes, resulting in a vector with significantly higher
in vitro
infection rates across a variety
of cells and tissues.
Recombinant AAV-2 and AAV-DJ vectors can be purified by CsCl gradient ultracentrifugation,
iodixanol discontinuous gradient ultracentrifugation, or high-performance liquid chromatography
(HPLC). The most popular technique, CsCl ultracentrifugation, is time consuming process which may
result in poor recovery and quality (nonviral protein contamination and a high ratio of genome copies
versus infectious units).
ViraBind™ AAV Purification Kit does not involve ultracentrifugation. AAV-2 or AAV-DJ is first
purified from viral supernatant with a single-step purification matrix, then further purified/concentrated
with a centrifugal concentrator (see Assay Principle below). The entire procedure takes about 3 hours.
Each preparation can handle one 150 mm dish or two 100 mm dishes
, resulting in 100 μL of highly
purified AAV-2 (Figure 1) or AAV-DJ with low ratio of genome copies versus infectious units.
ViraBind™ AAV Purification Kit provides an efficient system for quick purification of AAV-2 or
AAV-DJ with high recovery (>60%). The highly purified and concentrated viruses can be used in
primary cell infections and
in vivo
applications.
Summary of Contents for ViraBind Series
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