Engineering an Ionic Aggregation-Induced Luminescence-Labeled Fluorescence Lateral Flow Immunoassay for C-Reactive Protein in Human Plasma

Abstract

The surge of lateral flow immunoassays (LFAs) stimulates researchers to explore the novel vibrant aggregation-induced emission luminogen (AIEgen)-doped nanoparticles to improve the accuracy and reliability of LFAs. However, the loading amount of AIEgens currently used for the LFA in microspheres is limited due to their symmetrical large conjugated skeleton structure, which significantly reduces the fluorescence brightness of the signal reporter in the LFA. Herein, an ionic AIEgens with a donor–acceptor type was developed as the signal reporter of the LFA for C-reactive protein (CRP). Ionic AIEgens are unable to enter the hydrophobic cavity of polystyrene nanoparticles (PS) because of their low hydrophobic nature. By altering ionic AIEgens with extended alkyl chains, it is possible to increase their hydrophobicity, thereby potentially increasing the loading capacity within PS. Notably, the fluorescent nanoparticles (denoted as AIE_TPANPs) formed by embedding (E)-4-(4-(diphenylamino)styryl)-1-octadecylpyridin-1-ium iodide (TPA) in PS showed orange-red fluorescence emission and have high fluorescence quantum yield. Anti-CRP antibody (mAb1) could be effectively conjugated to the surface of AIE_TPANPs by an amino-carboxyl reaction, resulting in AIE_TPANPs-mAb1. The AIE_TPANPs-mAb1 exhibited a fluorescence emission at 613 nm, a point detectable by the naked eye with minimal background interference. The entire analysis was accomplished in just 10 min, achieving a limit of detection of 4.06 ng/mL for CRP. The AIE_TPANPs-mAb1-based LFA demonstrates excellent stability and specificity and fully meets the requirements for clinical diagnosis.