Biology Reference
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Fig. 1
AAV6 vector titration and quality control. (
a
) Measurement of the number of vector genome copies per
ml of vector suspension by qPCR. (
b
) Vector infectivity can be determined by fl ow cytometry, when the vector
encodes a fl uorescent protein. (
c
) Relative vector infectivity can be measured by qPCR following incubation of
the vector with cells permissive to infection. A reference vector of known effi ciency is used to establish a
standard curve to which the vector of unknown effi ciency is compared. (
d
) Quality of vector preparation is
assessed by separating denatured viral particles on a SDS-polyacrylamide gel. Pure vector suspensions should
only display three bands that correspond to each of the capsid proteins (VP1, 2, and 3)
across various vector sources [
48
]. Here, we provide a detailed
methodological approach to establish the titer of AAV6 vector
preparations. It is recommended to measure both the viral genome
content and the infectivity of vector batches to predict their effi -
cacy in vivo.
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