Fig. 2.

Establishment of the CLEIA system with FMU-CRT-17 mAb as coating antibody and HRP-conjucted FMU-CRT-8 mAb as detection antibody to detect soluble CRT. A Each of the six mAbs was used as coating antibody or as HRP-labeled detection antibody. The optical density (OD) 450 nm absorbance was detected for each antibody combination using sCRT concentration at 100 ng/ml. Histograms represented means of triplicate determinations (bars show mean with SD). B Standard calibration curves of sandwich ELISA with three combinations of anti-sCRT mAbs. Plotted values were obtained with 6.25, 12.5, 25, 50, 100, and 200 ng/ml rCRT by sandwich ELISA as described in materials and methods. Line graph is means of triplicate determinations. C Standard calibration curves of sCRT CLEIA with FMU-CRT-17 as coating mAb and HRP-labeled FMU-CRT-8 as detection mAb. Plotted values were obtained with 0.19, 0.39, 0.78, 1.56, 3.125, 6.25, 12.5, 25, 50, 100, and 200 ng/ml rCRT. The inset was the amplification of 0.19–3.125 ng/ml rCRT