First, Raman analysis was conducted to verify the covalent binding between antibody and polydiacetylene. Data wereprocessed to remove the effect of fluorescence by subtracting anappropriate baseline and normalized according to the measuredparameters. In the standardization process, the most intense peakof the “PDA” spectrum was adopted as a unit. Raman spectra for PDA vesicles and PDA/antibody vesicles show the characteristicpeaks due to the C C and CC backbone stretching vibrationsbetween 1400 cm−1and 1500 cm−1, 2000 cm−1and 2100 cm−1,respectively, and a complex, weaker structure below 1400 cm−1according to Bloor (1999). The C C and CC backbone wascaused by topopolymerization of diacetylene monomers in theorganized structures initiated by UV light irradiation yielding PDAthat constitutes alternating double and triple bonds (ene–eyne)along its backbone [9,12]. However, for PDA/antibody vesicles thepeak intensity in 1400 cm−1and 1500 cm−1was lower than PDAvesicles. There are two hypotheses for it: first, it could indicateantibody incorporation because there was a lower peak at thesewavenumbers; and second, it could be difficult for PDA vesiclesto polymerize in antibody presence as showed in Fig. 2 [21,25].However, from colorimetric perspective, the blue color intensitywas associated with the vesicle polymerization. So both vesi-cles, without and with antibody incorporation, showed the same absorbance intensity at 640 nm (blue color) indicating that the antibody was incorporated. Samples of PDA/antibody vesicles havespectra with less components in the C C and CC peaks and fewer, stronger peaks below 1400 cm−1than PDA vesicles. The peaks below 1400 cm−1 weaken and have a more complex structure than that of PDA/antibody vesicles [21,25].After this evidence of antibody incorporation in PDA vesicles,experiments were planned to obtain a quadratic model. First