eBook: Analytical Method and QC Testing Strategies for Biologics Production - 24

" As the gene therapy space matures, there will be more
demand for volume, higher dosage, and systems that
offer higher scalability, " Yan says. " With HEK293, the
maximum volume achieved in bulk would be around
200 liters, whereas with the Sf9/Baculovirus system,
manufacturers can comfortably get up to a thousand
liters. " Aside from the higher scalability, the Sf9/
Baculovirus expression system enables a high capacity
for multiple genes or a large insert (because of the large
and flexible viral genome), and biosafety (because
baculoviruses naturally do not infect humans).2
A quantitative real-time PCR
solution for residual genomic
Sf9 and Baculovirus DNA
Due to a lack of commercially-available assays for
residual DNA testing of Sf9 and Baculovirus DNA, drug
developers often need to build their own in-house
testing solutions. " These in-house tests require quite
a bit of process development, " says Dr. Kara Norman,
Senior Manager, R&D, Pharmaceutical Analytics at
Thermo Fisher Scientific. " For example, establishing
and maintaining a robust calibration standard requires
a lot of expertise and it's very time-consuming. " Norman
adds that strong inventory management and quality
control of individual components is also needed to
ensure lot-to-lot reproducibility.
To address these challenges, Thermo Fisher has
developed the first commercially-available duplex
qPCR assay for detecting Sf9 host cell DNA and
baculovirus DNA: resDNASEQ Quantitative Sf9 and
Baculovirus DNA Kit. The assay specifically targets two
genes: 1) fucosyl transferase on the Sf9 genome and 2)
Gp64 on the baculovirus genome. These amplicons are
both under 100 nucleotides, ensuring highly sensitive
detection of residual DNA. The assay is uniquely
designed to provide the following advantages in
testing:
(1) An integrated solution with a rapid, streamlined
workflow
assay functions as an end-to-end solution
including genomic standards and reagents, a sample
prep kit (PrepSEQ) for both automated and manual
nucleic acid extraction, Applied Biosystems TaqMan
real-time PCR technology, and data analysis software
(AccuSEQ). (Figure 1) The time taken from samples to
results is less than five hours.
This
Figure 1. The resDNASEQ solution is a flexible, integrated, end-to-end platform developed to help meet cGMP requirements.
24

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