Bifunctional ligand-mediated amplification of polydiacetylene response to biorecognition of diethylstilbestrol for on-site smartphone detection

Polydiacetylene (PDA) is very suited for sensitively detecting large biomolecules, and its unique chromatic properties enable visual read-out. However, application to the selective detection of small molecules remains challenging. Here, bifunctional ligands are studied to amplify the color change of PDA for biorecognition of small molecules for the smartphone-based detection of diethylstilbestrol (DES). PDA is decorated with streptavidin (PDA-SA, blue), and biotin-modified DES (bio-DES) is prepared as a bifunctional ligand to couple with PDA-SA and DES antibody. Since multiple bio-DES can bind to a single SA, then multiple SAs on PDA lead to an increased surface coverage of the vesicle. In samples without DES, PDA-SA-bio-DES-DES antibody complexes will form, leading to a color transition (blue to red); this color transition is greatly amplified by antibody-induced aggregation of the complexes. When DES is present, aggregation is inhibited due to competition for the antibody and PDA-SA-bio-DES retains its blue color. A linear relationship (0.4-1250 ng mL-1) is found between the colorimetric response and the logarithmic DES concentration, with adequate selectivity, accuracy (82.24-118.64%), and precision (below 8.24%). Finally, a paper-based DES PDA biosensor is developed with visual and smartphone-based detection limits of 10 ng mL-1 and 0.85 ng mL-1 in water, respectively.