eBook: Analytical Method and QC Testing Strategies for Biologics Production - 26

(2) Sensitivity: To evaluate sensitivity, a sample at the
limit of detection (LOD) was compared to the notemplate
control (NTC) by noting the cycle threshold
(Ct) value, or the number of PCR cycles required for the
fluorescent signal to cross the threshold (i.e. exceed
background level). The higher the Ct, the weaker the
reaction, and the lower the DNA concentration.
In all of the tests, the Ct value for the NTC was higher
than that of the LOD sample (30 fg). This clean
separation between the LOD and the NTC helps enable
customers to meet the regulatory requirements for
measuring extremely low levels of DNA. (Figure 3)
(3) Specificity: A variety of DNA types was tested to
ensure that the Sf9/Bac assays do not amplify and
produce a signal for off-target DNA. Samples were
spiked with 3ng of non-specific DNA, but no signal
was observed - all the cross reactants yielded a
negative result of high Ct (Ct > Ct of LOD). Both Sf9
and Baculovirus assays demonstrate high specificity
in the presence of genomic DNA from other sources.
(Figure 3)
(4) Singleplex vs multiplex performance: For this study,
singleplex Sf9 and singleplex baculovirus standard
curves were evaluated in the same conditions as
the combination standard curves. Results show the
standard curves overlapping, indicating the Sf9 and
Baculovirus assays do not interfere with each other,
and each target can be reliably measured on its own.
(Figure 4)
Figure 2. Standard curve performance across multiple sites, days, and operators on the 7500 Fast instrument. No significant difference
was found across all points on the standard curve. High precision, accuracy, linearity, and efficiency were achieved to enable quantitative
results across a broad range of DNA concentrations. (NTC = no-template control, IPC = internal positive control)
26

eBook: Analytical Method and QC Testing Strategies for Biologics Production

Table of Contents for the Digital Edition of eBook: Analytical Method and QC Testing Strategies for Biologics Production

eBook: Analytical Method and QC Testing Strategies for Biologics Production - 1
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 2
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 3
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 4
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 5
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 6
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 7
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 8
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 9
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 10
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 11
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 12
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 13
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 14
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 15
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 16
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 17
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 18
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 19
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 20
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 21
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 22
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 23
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 24
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 25
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 26
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 27
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 28
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 29
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 30
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 31
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 32
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 33
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 34
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 35
eBook: Analytical Method and QC Testing Strategies for Biologics Production - 36
https://www.nxtbookmedia.com