2 research outputs found
Single-Step Homogeneous Immunoassays Utilizing Epitope-Tagged Gold Nanoparticles: On the Mechanism, Feasibility, and Limitations
A single-step gold nanoparticle (AuNP)-based
immunoassay is demonstrated
in which the nanoparticle surface is tagged with short viral peptide
epitopes. Antiviral antibodies with monoclonal specificity trigger
nanoparticle aggregation yielding a colorimetric response that enables
detection of antibodies in the low-nanomolar range within a few minutes. <i>In silico</i> insights into the interactions at the epitope–gold
interface demonstrate that the conformational landscape exhibited
by the epitopes is strongly influenced by the amino acid sequence
and location of particular residues within the peptides. The conformation,
orientation, and linker chemistry of the peptides affect the immune
complex formation in nonintuitive ways that are, nevertheless, explained
by a unique sterically kinetically driven aggregation mechanism. The
rapid and specific performance of the AuNP immunoassay may have generic
potential in point of care diagnostics and other laboratory routines
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Strain-promoted cycloadditions in lipid bilayers triggered by liposome fusion
Due to the variety of roles served by the cell membrane, its composition and structure are complex, making it difficult to study. Bioorthogonal reactions, such as the strain promoted azide-alkyne cycloaddition (SPAAC), are powerful tools for exploring the function of biomolecules in their native environment but have been largely unexplored within the context of lipid bilayers. Here, we developed a new approach to study the SPAAC reaction in liposomal membranes using azide- and strained alkyne-functionalized Förster resonance energy transfer (FRET) dye pairs. This study represents the first characterization of the SPAAC reaction between diffusing molecules inside liposomal membranes. Potential applications of this work include in situ bioorthogonal labeling of membrane proteins, improved understanding of membrane dynamics and fluidity, and the generation of new probes for biosensing assays.</p