Biomedical Engineering Reference
In-Depth Information
TABLE 11.1 Assay Performance Characteristics Between the Two Validated PK
Methods for Drug Y
Validation Parameter (Acceptance
Criteria)
Flow Cytometric Pk
Assay
ECL-Based PK Assay
Specificity: Method can detect a
dose-dependent response
Pass
Pass
Accuracy: Assessed at 3-5 drug
concentrations with data collected
from 2-3 analysts (80-120%
recovery of the theoretical value)
80-120% recovery
Pass
80-120% recovery
Pass
Precision: Assessed at 3-5 concentra-
tions with data collected from 2-3
analysts (
<
15%CV for intra-assay;
<
20%CV for interassay)
Intra-assay (n
¼
5):
<
10%CV
Intra-assay (n
¼
6):
<
8%CV
Interassay (n
¼
4):
<
16%CV
Interassay (n
¼
5):
<
13%CV
Pass
Pass
LOD: Unspiked serum background
MFI
58.6 ng/mL
3.5 ng/mL
þ
3SD
LOQ: The lowest drug concentration
with acceptable accuracy (80-120%
recovery) and precision (
<
20%CV)
300 ng/mL
60 ng/mL
Linear: r
2
Linear: r
2
Linearity of sample dilution:
>
0.99 and
slope 0.95-1.09
>
0.99 and
slope 1.31-1.33
Additional parameters examined:
Curve-fit, sample
stability, normal sera
distribution, and
control ranges: Pass
Curve-fit, selectivity,
spike/recovery,
sample stability,
normal sera distri-
bution, and control
ranges: Pass
11.2.7.7 Provisional Control Reference Ranges
Data collected fromH, M, and L
QC samples in development studiesmay be used to establish provisional control ranges
for assay pass/fail criteria in validation studies. Upon completion of the validation
study, all QC data (from multiple interday runs by multiple analysts) should be
analyzed for establishment of control reference ranges for in-study/clinical sample
testing. A statistical analysis-based approach, such as mean
2SD, should be
considered to establish ranges. If the data set for each drug level is not normally
distributed, transformationmay be required prior to establishing the appropriate range.
11.3 ASSAY VALIDATION
Technical resources, such as FDA, EMEA, and ICH [3-6] guidelines on analytical
method validation, are available and should be referenced for flow cytometric method
validation studies. Additional details can be found in a paper published by DeSilva
et al. [7].
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