Inter- and intra-assay variation for the C3C assay.

<p>As quality controls (QC1-5), human sera was used. The controls (CO1 and -2) included in every run of the C3C ELISA were also included. The variation was calculated as the mean variation between 10 individual runs of each sample run in double determination.</p

Patient demographics.

<p>Patient demographics.</p

Percentage dilution recovery for the C3C assay using human serum and plasma samples.

<p>Samples were measured from undiluted to 1:4 diluted and linearity was assessed.</p

Spiking recovery of standard peptide in human serum, and high serum in low serum.

<p>The recovery (RE%) was calculated as the percentage recovery of the measured amount in the sample alone. The experiment was performed for three separate healthy human control sera. The standard peptide was added in 2-fold dilutions starting from 50 ng/mL (StdB) and high serum was added in 2-fold dilution starting from 1:2.</p

Products used for cleavage analysis.

<p>Products used for cleavage analysis.</p

Analyte stability in human serum.

<p>The serum samples were either subjected to four freeze/thaw cycles or stored at 4 or 20°C for 0, 2, 4 and 24 hours. All data are shown as mean percent recovery (RE%) compared to baseline (ie, 1 freeze/thaw cycle and storage time 0 hours, respectively).</p

Sequence alignment between the alpha-1 chain of type III collagen in humans, rats and mice.

<p>The antibody recognizes the neo-epitope starting from residue 642 to 651 in the human protein (ie, GLPGTGGPPG). 90% homology was observed for the human and mouse sequence due to an amino acid change at the second residue in the target sequence (L → I). 80% sequence homology was observed between the human and rat sequence due to two amino acid changes at the second (L → I) and sixth (G → S) position of the target sequence.</p

Generation of C3C by cleavage analysis of type III collagen.

<p>No reactivity was seen for either of the buffers or intact type III collagen (COL3). The only proteases generating C3C were the cathepsins B, L, S and K. Data are presented as mean ± SEM. Results below the detection limit were given the value of LLOQ. Statistical significance was determined by ANOVA with multiple comparison testing using the Tukey test with type III collagen, without any protease present (COL3), as a reference; ****p<0.0001.</p

Biomarker COL-18N correlate with ABR.

<p>Serum from 35 male HF patients aged 26 and over was measured with the COL-18N ELISA. Correlations between vascular endothelial type XVIII collagen concentration and ABR were analyzed using Spearman rank correlation coefficient and shown r = 0.45, p<0.006. Differences between ABR and COL-18N levels were considered statistically significant if p<0.05 and significant levels are displayed as: * = p<0.05; ** = p<0.01, and *** = p<0.001.</p

Dilution recovery of COL-18N assay.

<p>Dilution recovery of COL-18N assay.</p