The Experts below are selected from a list of 1926 Experts worldwide ranked by ideXlab platform
Dae-ro Ahn - One of the best experts on this subject based on the ideXlab platform.
-
A Microwell Plate-based multiplex immunoassay for simultaneous quantitation of antibodies to infectious viruses.
The Analyst, 2015Co-Authors: Min-suk Jeong, Dae-ro AhnAbstract:Antibodies (Abs) to disease-causing viruses in human blood are important indicators of infection status. While ELISA has been widely used to detect these Abs, a multiplex assay system for simultaneous detection of multiple Abs is still a desirable alternative method for a more efficient screening process because of the lack of multiplexing ability in ELISA. However, as all antibodies are based on immunoglobulin and recognized commonly by the same secondary antibody, it is impossible to multiplex the conventional indirect ELISA in a 96-Microwell Plate-based platform. To overcome this hurdle, we designed an assay consisting of two steps: capturing target Abs by specific antigens on DNA-encoded gold nanoparticles; and quantifying the target Abs by producing RNase H-mediated detection signals based on the DNA and additional RNA probes. With this newly designed method, we could simultaneously analyze three infectious disease-related Abs, anti-HIV Ab, anti-HCV Ab, and anti-HBV Ab, on the Microwell-based platform. The assay performance was evaluated by comparison with ELISA. Furthermore, the accuracy and precision of the assay in a practical application was also estimated by determining the amount of target Abs in human serum solutions.
-
elongated oligonucleotide linked immunosorbent assay for sensitive detection of a biomarker in a Microwell Plate based platform
Biosensors and Bioelectronics, 2013Co-Authors: Kicheol Han, Dae-ro Ahn, Eun Gyeong YangAbstract:Abstract The immunoassay is a representative method for detecting disease biomarkers and pathogenic biological agents. While the conventional enzyme-linked immunosorbent assay (ELISA) has been routinely used for the analysis of biological samples, methods with higher sensitivity are still in demand because the detection of low-level biomarkers is important for early diagnosis of lethal diseases. In this study, we developed a sensitive immunoassay called elongated oligonucleotide-linked immunoassay (EOLISA), employing long DNA oligonucleotides (80-mer), a fluorogenic RNA probe and RNase H for signal amplification. The elongated DNA oligonucleotides led to a highly amplified fluorescence signal via iterative cycles of DNA–RNA duplexation and subsequent degradation of the RNA in the duplex by RNase H. The immunoassay was evaluated for sensitive detection of fatty acid binding protein (FABP) in the 0–1 ng mL −1 range. When compared with ELISA, EOLISA showed about 10-fold improved detection sensitivity. With its simple procedure and reliable detection performance in the conventional platform, the proposed immunoassay is expected to have potential applications in clinical diagnostics.
Vladimir I. Razinkov - One of the best experts on this subject based on the ideXlab platform.
-
High-Throughput Analytical Light Scattering for Protein Quality Control and Characterization
Methods in molecular biology (Clifton N.J.), 2019Co-Authors: Daniel Some, Vladimir I. RazinkovAbstract:We present a review of high-throughput techniques for the characterization and quality control of proteins in the course of purification, evaluation, and formulation, based on static and dynamic light scattering. Multi-angle static light scattering (MALS) in combination with rapid, low-volume UHPLC size exclusion chromatography is effective in characterizing key biophysical properties, while dynamic light scattering (DLS) in high-throughput Microwell-Plate format provides large quantities of data in a short time to screen many conditions, excipients, cell lines, or candidate biotherapeutics.
Iva R Kennedy - One of the best experts on this subject based on the ideXlab platform.
-
Rapid determination of fumonisin B1 in food samples by enzyme-linked immunosorbent assay and colloidal gold immunoassay
Journal of agricultural and food chemistry, 2006Co-Authors: Shuo Wang, Nanju Lee, Ying Quan, Iva R KennedyAbstract:A rapid enzyme-linked immunosorbent assay (ELISA) test (Microwell Plate) and a membrane-based colloidal gold immunoassay in flow-through and lateral-flow formats for the rapid detection of fumonisin B1 (FB1) were developed. The rapid Microwell assay can be completed within 20 min with the detection limit of 0.5 ± 0.2 μg/L. Membrane-based colloidal gold immunoassays had a visual detection limit of 1.0 μg/L for FB1 with the detection time of
-
rapid determination of fumonisin b1 in food samples by enzyme linked immunosorbent assay and colloidal gold immunoassay
Journal of Agricultural and Food Chemistry, 2006Co-Authors: Shuo Wang, Ying Qua, Nanju Lee, Iva R KennedyAbstract:A rapid enzyme-linked immunosorbent assay (ELISA) test (Microwell Plate) and a membrane-based colloidal gold immunoassay in flow-through and lateral-flow formats for the rapid detection of fumonisin B1 (FB1) were developed. The rapid Microwell assay can be completed within 20 min with the detection limit of 0.5 ± 0.2 μg/L. Membrane-based colloidal gold immunoassays had a visual detection limit of 1.0 μg/L for FB1 with the detection time of <10 min. Matrix interference was eliminated by 15-fold dilutions of methanol extracts with buffer. These immunoassays can be used as quantitative or qualitative tools for the rapid detection of FB1 residues in 10−20 min on-site. Keywords: Fumonisin B1; rapid Microwell Plate ELISA; colloidal gold immunoassay; on-site screening
Michael J. Loeffelholz - One of the best experts on this subject based on the ideXlab platform.
-
gel electrophoresis southern blot and colorimetric Microwell Plate based system
2013Co-Authors: Michael J. Loeffelholz, Jiang FanAbstract:Infectious disease-related illnesses are a significant threat to human health resulting in substantial morbidity and mortality, worldwide. Timely and accurate diagnostic tools are critical for patient treatment decisions and disease outcomes. Molecular diagnostics are revolutionizing the clinical practice of infectious disease. The various formats of nucleic acid amplification are the most frequently used molecular tests in the diagnosis of infectious diseases due to its exquisite sensitivity and specificity. Gel electrophoresis and Southern hybridization are two basic technologies that are used to display the specific amplification of targeted gene and are still used in the laboratories for diagnosis because it is such a powerful technique, and yet reasonably easy and inexpensive.
-
Microwell Plate Detection Systems for Amplicon Detection and Characterization
Molecular Microbiology, 2011Co-Authors: Jiang Fan, Michael J. LoeffelholzAbstract:Sequence-specific methods for the detection of the products of nucleic acid amplification procedures (amplicons) have been developed for a variety of solid phases, including nylon membranes, Microwell Plates, microparticles, and, most recently, microchips (oligonucleotide probe microarrays). This chapter deals with procedures in which the molecules that capture amplicons are immobilized onto the surface of wells of Microwell Plates. Microwell Plate detection systems can be divided into two formats based on the molecule used to capture amplicons: an oligonucleotide probe (sequence-specific capture) and avidin (nonspecific capture). Microwell Plate detection systems have been developed in-house and are available commercially. Microwell Plate detection procedures are frequently based on capture and detection of biotinylated amplicons. This requires that the primer used in the PCR amplification to generate the amplicon strand complementary to the probe be tagged with biotin. The biotin substituent will allow the amplicon to be either captured or detected by avidin (or streptavidin), depending on the detection format. Polystyrene Microwell Plates are typically used for amplicon detection because of their high DNA binding capabilities. There is substantial literature demonstrating the excellent analytical performance and clinical utility of Microwell Plate detection systems for PCR amplicon detection. Microwell Plate detection of amplicons is flexible, is compatible with virtually any target, and can easily detect multiple pathogens under common hybridization and wash conditions.
Daniel Some - One of the best experts on this subject based on the ideXlab platform.
-
High-Throughput Analytical Light Scattering for Protein Quality Control and Characterization
Methods in molecular biology (Clifton N.J.), 2019Co-Authors: Daniel Some, Vladimir I. RazinkovAbstract:We present a review of high-throughput techniques for the characterization and quality control of proteins in the course of purification, evaluation, and formulation, based on static and dynamic light scattering. Multi-angle static light scattering (MALS) in combination with rapid, low-volume UHPLC size exclusion chromatography is effective in characterizing key biophysical properties, while dynamic light scattering (DLS) in high-throughput Microwell-Plate format provides large quantities of data in a short time to screen many conditions, excipients, cell lines, or candidate biotherapeutics.
