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CASE STUDY 1: PK PROFILES ON MULTIPLE MATRICES FROM INDIVIDUAL MICE
Ove Jonsson and his colleagues at AstraZeneca decided to see just how much data they could get from a single mouse while supporting a project on the elimination of human α-synuclein in mice. To do this, they used a CMS method to sample mice six times without exceeding the allowed total blood sampling volume, and prepared blood, serum and plasma (Ref 3; see also ‘The Capillary Sampling Method’). They then analyzed the samples with immunoassays run on Gyrolab xP workstation to generate PK profiles for individual mice. Their work also involved the first reported analysis of whole blood on Gyrolab xP workstation, which has immediate value in simplifying sample handling since it removes an additional step – the preparation of plasma.
α-SYNUCLEIN AND ASSAY
The target molecule, α-synuclein is believed to be involved in neurotransmitter regulation and is found at high levels in red blood cells and platelets in healthy individuals. This distribution makes hemolysis a potential analytical issue, and the study of plasma and blood levels particularly valuable. Two assays were developed – one cross-reactive for mouse and human α-synuclein and the other specific for human α-synuclein (results reported here).
RESULTS
The test of matrix effects indicated that the assays could tolerate up to 20% of matrix (blood, plasma or serum), which meant that, after dilution 1:4 (v/v), all matrices could be run on the same CD with one calibration curve.
Acceptance criteria were met for calibration-, QC- and study samples and the research group could collect all 648 samples as planned and in a timely manner.
The human-specific α-synuclein assay could be used to show a dose-dependent difference in mean human α-synuclein concentrations in plasma and blood from mice directly after intravenous dosing (see figure).
100 AUTHORS’ CONCLUSIONS
• CMS enabled collection and handling of 4 µL blood, plasma and serum samples
• Full PK profiles could be prepared from individual mice
• Six samples taken in 3 matrices from 36 individual mice could be collected and analyzed
• CMS samples are well suited for nanoliter-scale immunoassays
10 1 0.1
02 46 Time (h)
Human α-synuclein concentrations in plasma and blood determined with assay designed for human α-synuclein following a single intravenous administration of 2 mg/ kg human α-synuclein (n = 2–5). (Redrawn from Fig 5b in Olsson et al Ref 3)
D0024402/A 3
Plasma 2 mg/kg Blood 2 mg/kg
α-synuclein concentration (µ/mL)
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