A5028020050- Extracellular vesicles in disease
- SARS-CoV-2 and COVID-19 Research
- SARS-CoV-2 detection and testing
- MicroRNA in disease regulation
- Cell Adhesion Molecules Research
- Biosensors and Analytical Detection
- 3D Printing in Biomedical Research
- COVID-19 Clinical Research Studies
- Single-cell and spatial transcriptomics
- Monoclonal and Polyclonal Antibodies Research
- Nanopore and Nanochannel Transport Studies
- Advanced Biosensing Techniques and Applications
- T-cell and B-cell Immunology
- Systemic Lupus Erythematosus Research
- RNA Interference and Gene Delivery
- Tissue Engineering and Regenerative Medicine
- RNA Research and Splicing
- Inflammasome and immune disorders
- COVID-19 diagnosis using AI
- Anodic Oxide Films and Nanostructures
- Poxvirus research and outbreaks
- Caveolin-1 and cellular processes
- Immunotherapy and Immune Responses
- Kawasaki Disease and Coronary Complications
- Characterization and Applications of Magnetic Nanoparticles
Inspire
2020-2023
Brigham and Women's Hospital
2020-2022
Harvard University
2019-2022
BACKGROUND. Weeks after SARS-CoV-2 infection or exposure, some children develop a severe, life-threatening illness called multisystem inflammatory syndrome in (MIS-C). Gastrointestinal (GI) symptoms are common patients with MIS-C, and severe hyperinflammatory response ensues potential for cardiac complications. The cause of MIS-C has not been identified to date.
Abstract Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected over 21 million people worldwide since August 16, 2020. Compared to PCR and serology tests, SARS-CoV-2 antigen assays are underdeveloped, despite their potential identify active infection monitor disease progression. Methods We used Single Molecule Array (Simoa) quantitatively detect spike, S1 subunit, nucleocapsid antigens in the plasma of patients with (COVID-19). studied from 64 who were COVID-19...
Lymphoid follicles (LFs) are responsible for generation of adaptive immune responses in secondary lymphoid organs and form ectopically during chronic inflammation. A human model ectopic LF formation will provide a tool to understand development an alternative non-human primates preclinical evaluation vaccines. Here, it is shown that primary blood B- T-lymphocytes autonomously assemble into LFs when cultured 3D extracellular matrix gel within one channel two-channel organ-on-a-chip...
Sensitive assays are essential for the accurate identification of individuals infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here, we report a multiplexed assay fluorescence-based detection seroconversion in from less than 1 µl blood, and as early day first positive nucleic acid test after symptom onset. The uses dye-encoded antigen-coated beads to quantify levels immunoglobulin G (IgG), IgM IgA antibodies against four SARS-CoV-2 antigens. A logistic regression...
Extracellular vesicles (EVs) are released by all cells into biofluids and hold great promise as reservoirs of disease biomarkers. One the main challenges in studying EVs is a lack methods to quantify that sensitive enough can differentiate from similarly sized lipoproteins protein aggregates. We demonstrate use ultrasensitive, single-molecule array (Simoa) assays for quantification using three widely expressed transmembrane proteins: tetraspanins CD9, CD63, CD81. Using Simoa measure these EV...
Extracellular vesicles (EVs) are released by all cells into biofluids such as plasma. The separation of EVs from highly abundant free proteins and similarly sized lipoproteins remains technically challenging. We developed a digital ELISA assay based on Single Molecule Array (Simoa) technology for ApoB-100, the protein component several lipoproteins. Combining this ApoB-100 with previously Simoa assays albumin three tetraspanin found (Ter-Ovanesyan, Norman et al., 2021), we were able to...
Tests for COVID-19 generally measure SARS-CoV-2 viral RNA from nasal swabs or antibodies against the virus blood. It has been shown, however, that both particles and those are present in saliva, which is more accessible than We methods highly sensitive measurements of same saliva sample. developed an efficient extraction method combined it with ultrasensitive antibody test based on single molecule array (Simoa) technology. apply our to patients who presented hospital symptoms, some whom...
Coronavirus disease 2019 (COVID-19) manifests with high clinical variability and warrants sensitive specific assays to analyze immune responses in infected vaccinated individuals. Using Single Molecule Arrays (Simoa), we developed an assay assess antibody neutralization sensitivity multiplexing capabilities based on antibody-mediated blockage of the ACE2-spike interaction. The does not require live viruses or cells can be performed a biosafety level 2 laboratory within two hours. We used...
ABSTRACT Lymphoid follicles (LFs) are responsible for generation of adaptive immune responses in secondary lymphoid organs and form ectopically during chronic inflammation. A human model LF formation would provide a tool to understand development an alternative non-human primate models preclinical evaluation vaccines. Here, we show that primary blood B- T-lymphocytes autonomously assemble into ectopic LFs when cultured three-dimensional (3D) extracellular matrix gel within organ-on-a-chip...
Abstract Neuron-derived extracellular vesicles (NDEVs) present a tremendous opportunity to learn about the biochemistry of brain cells in living patients. L1CAM is transmembrane protein expressed neurons that presumed be found on NDEVs human biofluids. Previous studies have used immuno-isolation from plasma isolate for neurodegenerative disease diagnostics. We developed panel ultrasensitive Single Molecule Array (Simoa) assays known EV markers, as well L1CAM, and applied it study EVs...
Abstract Coronavirus disease 2019 (COVID‐19) manifests with high clinical variability and warrants sensitive specific assays to analyze immune responses in infected vaccinated individuals. Using Single Molecule Arrays (Simoa), we developed an assay assess antibody neutralization sensitivity multiplexing capabilities based on antibody‐mediated blockage of the ACE2‐spike interaction. The does not require live viruses or cells can be performed a biosafety level 2 laboratory within two hours. We...
Abstract Tests for COVID-19 generally measure SARS-CoV2 viral RNA from nasal swabs or antibodies against the virus blood. It has been shown, however, that both particles and those are present in saliva, which is more accessible than We methods highly sensitive measurements of serology same saliva sample. developed an efficient extraction method combined it with ultrasensitive test based on Single Molecule Array (Simoa) technology. apply our to patients who presented hospital symptoms, some...
Abstract The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has infected millions of people worldwide. PCR tests are currently the gold standard for diagnosis current disease (COVID-19) and serology used to detect seroconversion in patients. However, there is a lack quantitative ultra-sensitive viral antigen COVID-19. Here we show that Single Molecule Array (Simoa) assays can quantitatively SARS-CoV-2 spike, S1 subunit, nucleocapsid antigens plasma COVID-19 Combined with Simoa...
Abstract Extracellular vesicles (EVs) are released by all cells into biofluids and hold great promise as reservoirs of disease biomarkers. One the main challenges in studying EVs is a lack methods to quantify that sensitive enough can differentiate from similarly sized lipoproteins protein aggregates. We demonstrate use ultrasensitive, single molecule array (Simoa) assays for quantification using three widely expressed transmembrane proteins: tetraspanins CD9, CD63, CD81. Using Simoa measure...
Extracellular vesicles (EVs) are released by all mammalian cells and thought to be important mediators of intercellular communication. There many methods for isolating EVs from cell culture media, but one the most commonly used continues purification based on ultracentrifugation. This approach has advantage allowing a large input volume media. Here, we provide detailed protocol differential
Abstract New vaccine candidates evaluated in animals can lead to unpredicted toxicities or poor efficacy human clinical trials due species-specific differences immune responses. To address this problem, we developed the ectopic lymphoid follicle (LF) chip containing primary cells, including autologous blood B and T lymphocytes antigen presenting cultured a three-dimensional extracellular matrix (ECM) gel within an organ-on-a-chip microfluidic device. Superfusion via parallel channel...
Abstract Extracellular vesicles (EVs) are released by all cells into biofluids such as plasma. The separation of EVs from highly abundant free proteins and similarly-sized lipoproteins remains technically challenging. We developed a digital ELISA assay based on Single Molecule Array (Simoa) technology for ApoB-100, the protein component several lipoproteins. Combining this ApoB-100 with previously Simoa assays albumin three tetraspanin found (Ter-Ovanesyan*, Norman* et al., 2021), we were...
We present a protocol for differentiation of human induced pluripotent stem (iPS) cells into neurons in large quantities vitro isolation extracellular vesicles (EVs). employ iNGN (induced Neurogenin) expressing the transcription factors Neurogenin1/2 under control doxycycline-inducible promoter. EVs from conditioned media these can be isolated by differential ultracentrifugation.
Extracellular vesicles (EVs) are released by all mammalian cells and thought to be involved in intercellular communication. Here, we provide a detailed protocol for analyzing the Tetraspanins CD9, CD63 CD81 western blotting from EV lysate. These three proteins widely expressed different cell types considered "markers" of EVs.
Extracellular Vesicles (EV) are reservoirs of biomarkers such as mRNA and post-translationally modified proteins, which may be use in diagnosing disease. However, major challenges still remain the isolation characterization EVs complex biofluids plasma cerebrospinal fluid. We developed a method for quantifying by measuring three commonly expressed EV transmembrane proteins (CD9, CD63 CD81). Due to low abundance biofluids, we used single molecule array (Simoa) technology order quantify these...
Organ-on-a-Chip A microfluidic organ-on-a-chip model of the human lymphoid follicle that recapitulates vaccination responses in vitro. Blood-derived cells self assemble into germinal centers a matrix gel lower channel when nutrient medium is flowed through upper channel. Antigen-specific antibodies and cytokines are secreted response to seasonal influenza mirror observed patients. More details can be found article number 2103241 by Donald E. Ingber co-workers.