Preparation of dyed polymer microspheres by a physical-chemical dual-binding method and their application in lateral flow immunoassay

Lateral flow immunoassay (LFIA) has shown great competitiveness in point-of-care testing due to its flexibility and simplicity. Dyed polymer microspheres are one of the most widely used marker particles for signal presentation as they are very convenient for visual interpretation, which is one of the most attractive features of LFIA. The color intensity, as the most critical factor, is directly related to the visual effect. In this work, a physical-chemical dual-binding strategy was proposed for the preparation of functionalized dyed microspheres. Bifunctional seed microspheres were synthesized by introducing 4-vinylbenzyl chloride (VBC) into the soap-free emulsion polymerization process, and the effective immobilization of dyes inside and on the surface of the polymer microspheres was achieved by covalent bonding and swelling methods. The microspheres were characterized by SEM, FT-IR spectroscopy and UV-vis spectroscopy. The results showed that the microspheres containing VBC were spherical with an average particle size of 300 nm. When the microspheres were prepared by adding 10 wt% VBC relative to the total monomer, the immobilization amount of 1-[[4-[(dimethylphenyl)azo]dimethyl phenyl]azo]-2-naphthol (Red-27) was increased to 180 mg g-1, which was 1.8 times that of the microspheres without VBC. The resulting nanomaterials were successfully used to establish a lateral flow immunoassay for the detection of COVID-19 virus N protein. The linear response concentration range was 2.64-87.84 COI, and the detection limit was 14.95 COI. A physical-chemical dual-binding strategy was developed to prepare high-dyed microspheres, which were further used for the qualitative detection of COVID-19 virus N protein in LFIA.