Subproject #1 Analysis of biological samples using Immunoaffinity capillary electrophoresis (ICE) Capillary electrophoresis combined with lased induced fluorescent (LIF) detection is an established method for analyzing biological samples. The small sample volumes, short analysis times and low reagent use are advantageous over more traditional methods such as HPLC. Derivitization of the capillary with antibodies allows for specific capture and detection of single or multiple analytes from a complex biological sample. ICE increases the limits of detection for CE without the need for pre-processing samples. Additionally this technique can be used on both a traditional CE machine as well as a micro-chip CE. ICE has been used to process samples from several collaborators on the NIH campus. Studies include the following: -Analysis of human plasma samples for cytokines indicated in kidney diseases -Tissue analysis from mice and rats from disease model studies looking at Alzheimers -Analysis of plasma samples for antigens indicated in HIV disease models -Analysis of tissue samples from rats given cytokine inhibition drugs Subproject #2 Multiplex ELSIA analysis of cell secretions Multiplex ELISA improves upon the traditional ELISA format. Rather than detecting a single antigen in a single sample, multiplex technology allows for the detection of up to 16 antigens from a single sample. This reduces the overall time and cost associated with ELISA and requires far less sample volume to produce more information. The multiplex format is customizable to allow researchers to investigate a unique array of proteins and peptides. Studies using multiplex ELISA include: -Analysis of cell secretion products from astrocytes stimulated with growth factors -Analysis of cell secretion products from macrophage stimulated with growth factors Subproject #3 Surface Plasmon Resonance Biacore 3000 detection system uses Surface Plasmon Resonance (SPR) to monitor the refractive index (RI) change as molecules interact at the sensor chip surface. SPR is the excitation of surface plasmons by light. The ligand is immobilized on the sensor chip surface, while the analyte is injected in continuous flow over the surface. If an interaction takes place, the surface concentration of the analyte is increased which results in a RI change. SPR technology enables label-free real-time detection and monitoring of biomolecular events and provides qualitative and quantitative information on specificity of binding between two molecules, affinity, and kinetics of the interaction. Experimental design is highly flexible and the technology can be applied to protein interaction with other proteins, nucleic acids, lipids, small molecules. Studies using surface Plasmon resonance include: -Immobilization of Neutravidin (NA), strep-tadded Adenilate Cyclase (CyA) capturing and binding of cAMP Receptor (CRP) protein to CyA -Immobilization of 1D4 antibodies, Rho-tagged peripheral cannabinoid receptor CB2 capturing and the binding of antibody fragments (nanobodies, NBs) to CB2

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National Institute of Biomedical Imaging and Bioengineering
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Liu, Yijing; Yang, Zhen; Huang, Xiaolin et al. (2018) Glutathione-Responsive Self-Assembled Magnetic Gold Nanowreath for Enhanced Tumor Imaging and Imaging-Guided Photothermal Therapy. ACS Nano 12:8129-8137
Liu, Yijing; Wang, Zhantong; Liu, Yi et al. (2017) Suppressing Nanoparticle-Mononuclear Phagocyte System Interactions of Two-Dimensional Gold Nanorings for Improved Tumor Accumulation and Photothermal Ablation of Tumors. ACS Nano 11:10539-10548
Czikora, Agnes; Kedei, Noemi; Kalish, Heather et al. (2017) Importance of the REM (Ras exchange) domain for membrane interactions by RasGRP3. Biochim Biophys Acta Biomembr 1859:2350-2360
Phillips, Terry M; Wellner, Edward; McMohan, Shane et al. (2016) Measurement of Inflammatory Chemokines in Micro-dissected Tissue Biopsy Samples by Chip-Based Immunoaffinity Capillary Electrophoresis. Methods Mol Biol 1466:121-36
Liu, Yijing; Yang, Xiangyu; Huang, Zhiqi et al. (2016) Magneto-Plasmonic Janus Vesicles for Magnetic Field-Enhanced Photoacoustic and Magnetic Resonance Imaging of Tumors. Angew Chem Int Ed Engl 55:15297-15300
Cizza, Giovanni; Piaggi, Paolo; Lucassen, Eliane A et al. (2013) Obstructive sleep apnea is a predictor of abnormal glucose metabolism in chronically sleep deprived obese adults. PLoS One 8:e65400
Mendonca, Mark; Kalish, Heather (2013) Cytokine analysis by immunoaffinity capillary electrophoresis. Methods Mol Biol 919:259-71
Locatelli-Hoops, Silvia C; Gorshkova, Inna; Gawrisch, Klaus et al. (2013) Expression, surface immobilization, and characterization of functional recombinant cannabinoid receptor CB2. Biochim Biophys Acta 1834:2045-56
Zhao, Huaying; Gorshkova, Inna I; Fu, Gregory L et al. (2013) A comparison of binding surfaces for SPR biosensing using an antibody-antigen system and affinity distribution analysis. Methods 59:328-35
Kalish, Heather; Phillips, Terry M (2012) Assessment of chemokine profiles in human skin biopsies by an immunoaffinity capillary electrophoresis chip. Methods 56:198-203