'Fit-for-purpose' validation of SearchLight multiplex ELISAs of angiogenesis for clinical trial use.
AuthorsBacken, Alison C
Mitchell, Claire L
Jayson, Gordon C
Ward, Timothy H
AffiliationCR-UK Translational Angiogenesis Group, Paterson Institute for Cancer Research, University of Manchester, Wilmslow Road, Manchester, M20 4BX, UK.
MetadataShow full item record
AbstractValidated assays of circulating biomarkers of angiogenesis to predict and determine the efficacy of vascular-targeted anticancer drugs would facilitate successful drug development. Multiple biomarker candidates exist and a multiplex approach was sought to minimise the requisite patient blood volume and to aid selection of those biomarkers with greatest potential clinical utility. Validation of the SearchLight multiplex ELISA platform comprising two multiplex assays of nine potential angiogenesis biomarkers was conducted (plex 1; VEGF R1 and R2, IL-8, KGF, PlGF; plex 2; PDGFbb, HGF, FGFb and VEGF). The study focused on instrument qualification, analyte specificity within the multiplex format, assay precision and reproducibility. No evidence was found within the multiplex that signals output from one analyte impinged on another or that antibody cross-reactivity occurred. Spike recovery for 5 between-experiment repeats was within +/-15% of input values for 7 of the 9 multiplexed analytes, with a coefficient of variation (CV) of <20% for 6 of the 9 analytes. Plasma samples from 8 ovarian cancer patients (who were not receiving therapy) were assessed using the two multiplexes on this platform to explore the likely baseline variability in this disease context. This study suggests that the platform and the multiplex approach will be useful to evaluate pharmacodynamic responses to vascular targeted therapy in early clinical trials.
Citation'Fit-for-purpose' validation of SearchLight multiplex ELISAs of angiogenesis for clinical trial use. 2009, 342 (1-2):106-14 J. Immunol. Methods
JournalJournal of Immunological Methods
- Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples.
- Authors: Bastarache JA, Koyama T, Wickersham NE, Ware LB
- Issue date: 2014 Jun
- Accuracy and reproducibility of a multiplex immunoassay platform: a validation study.
- Authors: Bastarache JA, Koyama T, Wickersham NE, Mitchell DB, Mernaugh RL, Ware LB
- Issue date: 2011 Mar 31
- Issues on fit-for-purpose validation of a panel of ELISAs for application as biomarkers in clinical trials of anti-Angiogenic drugs.
- Authors: Brookes K, Cummings J, Backen A, Greystoke A, Ward T, Jayson GC, Dive C
- Issue date: 2010 May 11
- Method validation and preliminary qualification of pharmacodynamic biomarkers employed to evaluate the clinical efficacy of an antisense compound (AEG35156) targeted to the X-linked inhibitor of apoptosis protein XIAP.
- Authors: Cummings J, Ranson M, Lacasse E, Ganganagari JR, St-Jean M, Jayson G, Durkin J, Dive C
- Issue date: 2006 Jul 3
- Simultaneous measurement of multiple ear proteins with multiplex ELISA assays.
- Authors: Trune DR, Larrain BE, Hausman FA, Kempton JB, MacArthur CJ
- Issue date: 2011 May