Whole blood was collected under CO2/O2 anesthesia
into tubes containing EDTA as an anticoagulant on study days 2, 7, 14, 21, 22,
45, and 60. Specimens were collected and preserved as single aliquots and
stored in a freezer set to maintain −70°C. Serum was collected in tubes containing serum separator gel. The cardiac safety
biomarkers analyzed were cTnI, cTnT, BNP, NT-proBNP, ANP and CK. The BNP,
NT-proBNP, and cTnI, and cTnT were analyzed using biomarker kits from Mesoscale
Diagnostics (catalog numbers of K153KFD, K153JKD, and K15161C respectively)
while ANP was analyzed using biomarker kit from Shanghai BlueGene Biotech
(catalog number of E02A0493). The BNP, NT-proBNP, cTnI, and cTnT assay plates
were read by the MesoScale Diagnostic’s Sector Imager 2400 model 1250
and the ANP assay plates were read by the μQuant plate reader using KC4
software. The data were then transferred to Softmax Pro 5 for generation of the
reference curves and sample analysis. All assay reference curves were qualified
to determine if the assay kits were performing as per the manufacturer
instructions. Each reference curve was analyzed a minimum of twelve times in
duplicate to demonstrate reproducibility.
into tubes containing EDTA as an anticoagulant on study days 2, 7, 14, 21, 22,
45, and 60. Specimens were collected and preserved as single aliquots and
stored in a freezer set to maintain −70°C. Serum was collected in tubes containing serum separator gel. The cardiac safety
biomarkers analyzed were cTnI, cTnT, BNP, NT-proBNP, ANP and CK. The BNP,
NT-proBNP, and cTnI, and cTnT were analyzed using biomarker kits from Mesoscale
Diagnostics (catalog numbers of K153KFD, K153JKD, and K15161C respectively)
while ANP was analyzed using biomarker kit from Shanghai BlueGene Biotech
(catalog number of E02A0493). The BNP, NT-proBNP, cTnI, and cTnT assay plates
were read by the MesoScale Diagnostic’s Sector Imager 2400 model 1250
and the ANP assay plates were read by the μQuant plate reader using KC4
software. The data were then transferred to Softmax Pro 5 for generation of the
reference curves and sample analysis. All assay reference curves were qualified
to determine if the assay kits were performing as per the manufacturer
instructions. Each reference curve was analyzed a minimum of twelve times in
duplicate to demonstrate reproducibility.