Fig. 5

Correlation between SAST and dSAST experimental values. a dSAST values of the eight clones obtained by the EW method were plotted on the vertical axis and the SAST values on the horizontal axis; an approximate equation between the two data sets was calculated, yielding a slope of 0.1988. This indicated that the dSAST values were lower than the SAST values; however, these mAbs had reactivity to the antigen after thermal denaturation. b dSAST and SAST values were plotted for the 54 clones obtained by the MW method as in (a). The approximate equation showed a slope of 0.107, which was smaller than the slope of EW, indicating that the reactivity between mAb and antigen was reduced owing to thermal denaturation. c dSAST and SAST values of 65 clones were obtained by MS combination as in (a), and the slope of the approximate equation was 0.0648. This indicates that the antigenicity recognized by MS antibodies was mostly lost after thermal denaturation. d ELISA and SAST values of 32 WB positive clones of MS were plotted, and an approximate equation was obtained. e ELISA and SAST values of 33 WB negative clones of MS were plotted, and an approximate equation was obtained. Note that the slope of the approximate equation of (d) was slightly higher than that of (e); however, although there was no significant difference between the two values. f Using Student's t-test, we examined the significance between the slopes of the three plots; no statistically significant difference was observed between EW and MW; however, the slope was slightly lower for MS. A significant difference was observed between EW and MS, indicating that mAbs recognizing epitopes sensitive to thermal denaturation were obtained in MS compared to EW. ***: p < 0.01