Highly Sensitive Detection of Bacteria by Binder-Coupled Multifunctional Polymeric Dyes

Abstract

Infectious diseases caused by pathogens are a health burden, but traditional pathogen identification methods are complex and time-consuming. In this work, we have developed well-defined, multifunctional copolymers with rhodamine B dye synthesized by atom transfer radical polymerization (ATRP) using fully oxygen-tolerant photoredox/copper dual catalysis. ATRP enabled the efficient synthesis of copolymers with multiple fluorescent dyes from a biotin-functionalized initiator. Biotinylated dye copolymers were conjugated to antibody (Ab) or cell-wall binding domain (CBD), resulting in a highly fluorescent polymeric dye-binder complex. We showed that the unique combination of multifunctional polymeric dyes and strain-specific Ab or CBD exhibited both enhanced fluorescence and target selectivity for bioimaging of Staphylococcus aureus by flow cytometry and confocal microscopy. The ATRP-derived polymeric dyes have the potential as biosensors for the detection of target DNA, protein, or bacteria, as well as bioimaging.

Flow cytometry: Aspects and application in plant and biological science

“…The ability to direct cell labelling in vivo through injection into the bloodstream is the primary benefit of this method [50]. There are several fluorescent labels that may be conjugated with antibodies, including small-molecule organic dyes, fluorescent proteins, quantum dots, polymer dyes, and tandem dyes [51,52] (Table 1).…”

Tailored Branched Polymer–Protein Bioconjugates for Tunable Sieving Performance

“…The technique was extended to the grafting of well-controlled hyperbranched polymers directly from biomacromolecules. 41 Herein, using green-light-induced CRBP, 26,44,45 and a highthroughput synthetic setup, we developed a first straightforward approach to introduce branching into protein−polymer hybrids using inibramers. We prepared well-defined bioconjugates of proteins with branched polymers allowing for tunable degrees of branching in one-pot (Figure 1).…”

Hydrophilic Poly(meth)acrylates by Controlled Radical Branching Polymerization: Hyperbranching and Fragmentation

“…ATRP of acrylates in water is still considered a challenge due to potential hydrolysis of the C­(sp 3 )–X (secondary halides are more prone to hydrolysis than tertiary), leading to loss of chain-end functionality since the C­(sp 3 )–OH cannot participate in further chain growth. − Second, the high values of equilibrium constant of ATRP in water lead to a high concentration of radicals, which may result in more dead chains. , These challenges shall be even more pronounced during the controlled synthesis of hyperbranched acrylates using inibramers because their incorporation enhances the concentration of radicals. , Recently, we extended the scope EY/Cu photocatalyzed ATRP resulting in well-defined water-soluble linear acrylates under biorelevant conditions − using low-energy green light. , The use of Cu II /Me 6 TREN as deactivator and EY at ppm levels in 1× phosphate-buffered saline (PBS) medium suppressed the dissociation of the [X–Cu II /L] + deactivators and disproportionation of Cu I species to Cu II and Cu 0 thereby maintaining equilibrium throughout ATRP.…”

“… 63 , 64 Recently, we extended the scope EY/Cu photocatalyzed ATRP 65 resulting in well-defined water-soluble linear acrylates under biorelevant conditions 66 − 68 using low-energy green light. 69 , 70 The use of Cu II /Me 6 TREN as deactivator and EY at ppm levels in 1× phosphate-buffered saline (PBS) medium suppressed the dissociation of the [X–Cu II /L] + deactivators and disproportionation of Cu I species to Cu II and Cu 0 thereby maintaining equilibrium throughout ATRP.…”