Article Figures & Data
Figures
- Figure 1.
Functional flow chart of the PD assay development, implementation, and support strategy used in the Division of Cancer Treatment and Diagnosis for support of early-stage clinical trials. FFP, fit for purpose; SOP, standard operating procedure.
- Figure 2.
γ-H2AX detection in CTC. A, photomicrograph of γ-H2AX-postive MCF-7 breast cancer cells used as the positive control. Cells were treated with 1 μmol/L topotecan for 2 hours and stained with γ-H2AX-AF488 (green) and DAPI (blue). Image capture was conducted using Leica 5000 DM. Final magnification, ×200. B and C, images of γ-H2AX-postive CTCs from a breast cancer patient treated with ABT-888 combined with cytoxan. The blood samples were obtained from the patient in cycle one on day 2, 24 hours after drug administration (B) and day 5, 24 hours after drug administration (C). The EpCAM-enriched CTCs were stained with conjugated antibodies for CK-PE, CD45-APC, DAPI, and γ-H2AX-AF488 and analyzed on the CellSearch system.
Tables
- Table 1.
Multiplexa panels of apoptosis biomarkers
Panel 1 Panel 2 Panel 3 1. BAK, total 6. BIM, total 11. BAK–Mcl1 heterodimer 2. BAX, total 7. BAD, total 12. BAK–Bcl-(X)l heterodimer 3. Caspase-3, total 8. BAX–Bcl2 heterodimer 13. Caspase-3, cleaved (only) 4. Lamin-B 9. Bcl-(X)l, total 14. Phospho-S99BAD 5. Smac (dimer) 10. Mcl-1, total 15. Survivin -
↵aMultiplexing is achieved on a Luminex platform, which utilizes bead-based technology to multiplex up to 100 assays. The sandwich immunoassay uses a capture antibody covalently linked to beads and a reporter antibody reactive to separate epitope, linked to a fluorescent probe. In the case of heterodimers, the immobilized antibody captures one component and labeled reporter antibody binds to second component of the heterodimers. The validity of each measurement was demonstrated by immunoprecipitation and Western blot analysis of each biomarker in cancer cell or tumor tissue lysates.
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Volume 20, Issue 10
