The multiplexed detection of cell surface proteins plays an integral role in the differentiation of cell types and diagnosis of disease. Currently, the use of flow cytometry to evaluate fluorescently labelled surface proteins is the gold standard technique for the diagnosis of diseases such as chronic lymphocytic leukemia, the most prevalent blood cancer in the western world. However, the number of proteins that can be detected at once is limited by the broad emission signals and there is a push to create Raman-based labels whose narrow spectral signatures allow for greater vmultiplexing. Detection methods focusing on the coupling of the localized surface plasmon resonances (LSPRs) of plasmonic surfaces with Raman active dyes as a way to generate probes have been shown to be the most promising methods to address the brightness limitation. To be practical alternatives to fluorescent markers, these probes must: 1. Produce strong & detectable SERS signals, 2. Produce signals that are consistent over time & between batches and a quantitative technique; 3. Demonstrate colloidal stability in physiological media, and 4. Specifically target proteins with minimal non-specific binding. We will describe 3 particle types: PEGylated gold core NP, liposome encapsulated gold nanoparticles, and j-aggregate metal core np.<br/>The j-aggregate particles are especially bright. They exhibit an excitonic resonance that is key in setting up a strong electric field enhancement in the j-aggregate layer. The collective absorption of J-aggregates can tightly couple with LSPRs of NPs to result in greater SERS. In producing these, a cationic linker facilitates formation of J-aggregates on the NP surface. Silica encapsulation provides colloidal stability and brings dyes closer to surface to provide fluorescence quenching.<br/>Liposomes composed of zwitterionic lipids limit biofouling and serve as a modular matrix to incorporate a variety of functional molecules. Dark field microscopy and SERS represent combined functionalities for targeted liposomal probes. Two methods of antibody conjugation to SERS liposomes are reported: (i) direct conjugation to functional groups on the SERS liposome surface, or (ii) post-insertion of lipid-functionalized antibody fragments (Fabs) into preformed SERS liposomes. In vitro experiments targeting both the lymphoma cell line LY10 and primary human chronic lymphocytic leukemia (CLL) cells demonstrate the usefulness of these probes as optical contrast agents.

Nanoparticles for surface enhanced Raman scattering detection of cell surface proteins

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.