After isolating the anti-paclitaxel antibody 33D9, we opted to employ it as a capture antibody for any drug conjugated with paclitaxel, provided that the drug can be fluorescently labeled. To confirm its functionality, we conducted an assay using the 52B8hG1ptx antibody conjugated to paclitaxel and generated a standard curve. This assay utilized the double-antibody sandwich method in an enzyme-linked immunosorbent assay (ELISA) as depicted in Figure 1. Our findings indicate that, within a specific range, there's a direct correlation between the concentration of the antibody-drug conjugate and the resulting fluorescence intensity

 

The results of the experiment show that we obtained a standard curve that can show the relationship between the concentration of 52B8hG1ptx and the fluorescence value, where the statistical method utilized is the four-parameter fitting method. The R² value of the standard curve is 0.99, which indicates that the polynomial curve fits to the detection data very well, and is used to predict new data with an accuracy of up to ninety-nine percent.

 

DISCUSSION: As can be seen from the result graph above, this assay has a detection limit, after the concentration of 52B8hG1ptx exceeds 80ng/ml, its fluorescence value will no longer increase with the concentration, and there is no longer a linear relationship. When the concentration of 52B8hG1ptx is lower than 0.6ng/ml, its fluorescence value will not decrease with the decrease of concentration, and there is no longer a linear relationship. Therefore, the detection limit of this product in the detection of this drug: 0.6ng/ml-80ng/ml. the data are shown in the figure (Figure 3).

Figure 3 Raw data