Recent years have confirmed the ubiquitous applicability of lateral flow assays (LFA) in point-of-care testing (POCT). To make this technology available for low abundance analytes, strategies towards lower limits of detections (LOD), while maintaining the LFA’s ease of use, are still being sought. Here, we demonstrate how liposomes can significantly improve the LOD of traditional gold nanoparticle (AuNP)–based assays while fully supporting a ready-to-use system for commercial application. We fine-tuned liposomes towards photometric and fluorescence performance on the synthesis level and applied them in an established interleukin 6 (IL-6) immunoassay normally using commercial AuNP labels. IL-6’s low abundance (< 10 pg mL−1) and increasing relevance as prognostic marker for infections make it an ideal model analyte. It was found that liposomes with a high encapsulant load (150 mmol L−1 sulforhodamine B (SRB)) easily outperform AuNPs in photometric LFAs. Specifically, liposomes with 350 nm in diameter yield a lower LOD even in complex matrices such as human serum below the clinically relevant range (7 pg mL−1) beating AuNP by over an order of magnitude (81 pg mL−1). When dehydrated on the strip, liposomes maintained their signal performance for over a year even when stored at ambient temperature and indicate extraordinary stability of up to 8 years when stored as liquid. Whereas no LOD improvement was obtained by exploiting the liposomes’ fluorescence, an extraordinary gain in signal intensity was achieved upon lysis which is a promising feature for high-resolution and low-cost detection devices. Minimizing the procedural steps by inherently fluorescent liposomes, however, is not feasible. Finally, liposomes are ready for commercial applications as they are easy to mass-produce and can simply be substituted for the ubiquitously used AuNPs in the POCT market. Graphical abstract

Liposomes have been widely applied in bioanalytical assays. Most liposomes used bare negative charges to prevent non-specific binding and increase colloidal stability. Here, in contrast, highly stable, positively charged liposomes entrapping the fluorescent dye sulforhodamine B (SRB) were developed to serve as a secondary, non-specific label‚ and signal amplification tool in bioanalytical systems by exploiting their electrostatic interaction with negatively charged vesicles, surfaces, and microorganisms. The cationic liposomes were optimized for long-term stability (> 5 months) and high dye entrapment yield. Their capability as secondary, non-specific labels was first successfully proven through electrostatic interactions of cationic and anionic liposomes using dynamic light scattering, and then in a bioassay with fluorescence detection leading to an enhancement factor of 8.5 without any additional surface blocking steps. Moreover, the cationic liposomes bound efficiently to anionic magnetic beads were stable throughout magnetic separation procedures and could hence serve directly as labels in magnetic separation and purification strategies. Finally, the electrostatic interaction was exploited for the direct, simple, non-specific labeling of gram-negative bacteria. Isolated Escherichia coli cells were chosen as models and direct detection was demonstrated via fluorescent and chemiluminescent liposomes. Thus, these cationic liposomes can be used as generic labels for the development of ultrasensitive bioassays based on electrostatic interaction without the need for additional expensive recognition units like antibodies, where desired specificity is already afforded through other strategies.