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Immunoassay - ELISA Research

06:06 EST 20th February 2019 | BioPortfolio

Interference of High Dose Biotin Supplementation with Thyroid Parameters in Immunoassays Utilizing the Interaction between Streptavidin and Biotin: a Case Report and Review of Current Literature.

Automated immunoassays utilizing the interaction between streptavidin and biotin are widely used. Nonetheless, biotin remains an often overlooked confounder.

Microencapsulated Immunoassays for Detection of Cytokines in Human Blood.

Cytokines are produced by leukocytes in blood and may be used as indicators of malignancies or infections. The objective of this study was to develop a strategy for immunosensing cytokines in whole, unprocessed human blood. Microfluidic droplet generation was employed to fabricate ∼400 μm diameter microcapsules with a hydrogel shell and an aqueous core containing sensing microbeads. The hydroge...

The use of ErpY-like recombinant protein from Leptospira interrogans in the development of an immunodiagnostic test for swine leptospirosis.

Swine leptospirosis poses a major problem in the agricultural sector. The gold standard for serodiagnosis of leptospirosis is Microscopic Agglutination Test (MAT). However, the limitations of this technique make the search for alternative diagnostic methods inevitable. In the present study, ErpY-like recombinant protein (rErpY-like), produced in Escherichia coli and used as antigen in indirect enz...

MicroRNA-30c abrogation protects against spinal cord ischemia reperfusion injury through modulating SIRT1.

Spinal cord ischemia/reperfusion (I/R) injury is a severe complication in many surgeries. Although microRNAs (miRNAs) are involved in I/R-caused spinal cord injury (SCI), the mechanism that underlies miR-30c interacted with SCI remains elusive. In this study, I/R surgery or oxygen-glucose deprivation (OGD) were performed to establish SCI model in vivo or in vitro, respectively. Basso, Beattie and ...

Validation of LC-MS/MS methods for the determination of mirabegron and eight metabolites in human plasma in a paediatric population.

Mirabegron is the first registered β3-adrenoceptor agonist for treatment of overactive bladder as an alternative to antimuscarinics. Previously four liquid chromatography-tandem mass spectrometry assays were published to determine mirabegron and eight metabolites (M5, M8, M11-M16) in human plasma. In order to support paediatric development, the assays were further optimized to reduce the required...

Development of Caco-2 cells-based gene reporter assays and evaluation of herb-drug interactions involving CYP3A4 and CYP2D6 gene expression.

The indiscriminate use of medicinal plants and herbal medicinal products concomitantly with conventional drugs may result in herb-drug interactions that may lead to fluctuations in drug bioavailability, therapeutic failure, and/or toxic effects. CYP450 enzymes play an important role in drug biotransformation and herb-drug interactions. Thus, the aim of this study was to develop and apply Caco-2 ce...

General detection of Shiga toxin 2 and subtyping of Shiga toxin 1 and 2 in Escherichia coli using qPCR.

Shiga toxin-producing E. coli (STEC) is a gastrointestinal pathogen and has been recognized as one of the serious problems in public health. Shiga toxin genes (stx) can be grouped into different types according to their differences in sequence and biological activities. The two main groups of stx are stx and stx with each group containing several subtypes. It is essential to develop rapid stx/stx ...

scFv6.C4 DNA vaccine with fragment C of Tetanus toxin increases protective immunity against CEA-expressing tumor.

The carcinoembryonic antigen (CEA) is the main tumor-associated antigen of colorectal cancers. Previously, we developed a DNA vaccine using scFv6.C4, a CEA surrogate, against CEA-expressing tumors; 40% of the vaccinated mice were tumor-free after tumor challenge. In order to enhance vaccine efficacy, fragment C of Tetanus Toxin (FrC) was tested as adjuvant. C57BL/6J-CEA2682 mice were electroporate...

Development of a sandwich enzyme-linked immunosorbent assay for the quantification of ponatinib in serum.

The tyrosine kinase inhibitor ponatinib is extensively metabolized in the body, and consequently the development of specific immunoassays for pharmacokinetic studies and therapeutic drug monitoring of ponatinib is challenging. If two antibodies simultaneously recognize the entire structure of ponatinib, they could be utilized to establish an ultra-specific sandwich immunoassay for ponatinib, free ...

A comparative study of experimental and commercial serological tests for detection of antibodies in humans with tularemia.

Experimental serological tests were developed to determine anti-tularensis antibodies in humans in immunochromatography formats (LF-test LPS Ft15) and enzyme immunoassay (ELISA LPS Ft15) using as an antigen highly purified LPS F. tularensis 15 NIIEG. Analysis was conducted of the sensitivity and specificity of the developed tests and commercial tularemia antigen «RNGA-Tul-AG» (production Stavrop...

Application of Mass Spectrometry-Based Immunoassays for the Species- and Tissue-Specific Quantification of Banned Processed Animal Proteins in Feeds.

Processed Animal Proteins (PAPs) are considered as a sustainable protein source to improve the nutritional profile of feed for livestock and aquaculture. However, the use of these proteins is strongly regulated since the bovine spongiform encepha-lopathy (BSE) crisis. The reintroduction of non-ruminant PAPs for the use in aquaculture in 2013 has driven the need for alternative analytical methods t...

A Practical ELISA for Azaspiracids in Shellfish via Development of a New Plate-Coating Antigen.

Azaspiracids (AZAs) are a group of biotoxins that appear periodically in shellfish and can cause food poisoning in humans. Current methods for quantifying the regulated AZAs are restricted to LC-MS but are not well suited to detecting novel and unregulated AZAs. An ELISA method for total AZAs in shellfish was reported recently, but unfortunately, it used relatively large amounts of the AZA-1-conta...

Evaluation of procalcitonin immunoassay concordance near clinical decision points.

Background Procalcitonin (PCT) is a biomarker for systemic bacterial infections and may aid in decision making for antimicrobial stewardship. Numerous PCT assays are available on common clinical immunoassay platforms. However, questions remain about the harmonization of these assays and whether the same clinical decision points may be used with all methods. Methods Thirty-seven remnant patient ser...

Generation and characterization of D-dimer specific monoclonal antibodies for use in latex agglutination test.

The commercially available D-dimer assays used in the clinical practice often show differences in the results, and their specificity and sensitivity are rather unsatisfactory. Our aim was to develop a new monoclonal antibody against D-dimer with a proper specificity, and estimating its suitability using in a latex agglutination diagnostic test. Monoclonal antibodies were generated using hybridoma ...

Human chorionic gonadotropin suspected heterophile interference investigations in immunoassays: a recommended approach.

Background Heterophile antibody (HAb) interferences in immunoassays can cause falsely elevated hCG concentrations leading to incorrect diagnosis and treatments options. When results are not consistent with the clinical findings, hCG HAb interference investigation may be requested by the physician. A retrospective evaluation of the frequency of HAb interference was performed among cases of physicia...

A novel point-of-care oral anti-HCV assay: Is it reliable for screening hepatitis C virus infection in the era of direct-acting antivirals?

Recent advance in the direct-acting antivirals (DAAs) offers the potentials to eradicate hepatitis C virus (HCV) worldwide and makes universal screening more urgent. A point-of-care (POC) oral anti-HCV assay, the Fortune assay, was developed and its performance was evaluated. Individuals with or without HCV infection were recruited in three Centers. Paired oral and serum samples were tested using ...

Immunogenicity of Golimumab and its Clinical Relevance in Patients With Ulcerative Colitis.

Antidrug antibody (ADA) detection with standard bridging enzyme immunoassays (EIA) can yield false-negative results or underestimate titers through drug interference. A more sensitive assay was needed to determine clinical impact of antigolimumab antibodies.

A Novel Enzyme Immunoassay for the Detection of Buprenorphine, Norbuprenorphine and Their Glucuronides in Urine.

Buprenorphine is a commonly used opioid in pain therapy as well as in opiate maintenance therapy. Immunoassays are quick and cost-effective methods for the necessary toxicological urine analysis of maintenance therapy patients. In this study a novel enzymatic immunoassay, the Thermo Fisher Scientific CEDIA Buprenorphine II assay (Bup2) was evaluated for the detection of buprenorphine, norbuprenorp...

Synthesis of hapten, generation of specific polyclonal antibody and development of ELISA with high sensitivity for therapeutic monitoring of crizotinib.

Crizotinib (CZT) is a potent drug used for treatment of non-small cell lung cancer (NSCLC); however, its circulating concentration variability has been associated with acquired resistance and toxicity, restricting the success of cancer treatment. As such, the development of an assay that monitors CZT plasma concentrations in patients is a valuable tool in cancer treatment. In this study, a hapten ...

Multiplexing Protein and Gene Level Measurements on a Single Luminex Platform.

The ability to simultaneously measure multiple secreted proteins and the corresponding gene expression levels from a single sample is valuable for comprehensive analysis. Bottlenecks to traditional immunoassays and gene expression assays include large sample consumption, time consuming experimental procedures, and complex data analysis.

Evaluation of the Cutoff of Anti-HCV Antibody Enzyme-Linked Immunosorbent Assay in 7 Blood Station Laboratories.

To evaluate the necessity and suitability of the anti-HCV ELISA teot gray zone setted up by 7 blood station laboratories.

Evaluation of the Abbott ARCHITECT™ cytomegalovirus IgM/IgG, rubella IgM/IgG, and syphilis treponemal antibodies enzyme immunoassays in a mother and child health center population.

This study evaluated the concordance of Architect™ chemiluminescent microparticle immunoassays with Captia™ ELISA for cytomegalovirus (CMV) IgM and IgG, with Enzygnost™ and Captia™ ELISA for rubella IgM and IgG and with Trep-Sure™ ELISA for syphilis treponemal antibodies in a mixed pediatric and obstetrical population. Total agreement between assays and Kappa statistic value were 82.5% (...

A new high-resolution melting analysis for the detection and identification of Plasmodium in human and Anopheles vectors of malaria.

Among vector-borne diseases malaria is the leading cause of morbidity in the world, with more than 200 million cases per year and a large number of deaths. The techniques traditionally used for the detection of Plasmodium in humans and Anopheles mosquitoes include microscopy, IRMA, ELISA, antibody or molecular assays, and anopheline dissection. However, these techniques are limited by their requir...

Identifying Protein-DNA and Protein-Protein Interactions in Avian Embryos.

The chick embryo is a powerful model for experimental embryology due to its accessibility, sturdiness, and ease of manipulation. Here we describe protocols for analysis of protein-DNA and protein-protein interactions in tissues and cells isolated from the developing chick. These assays are aimed at the identification of interactions between transcription factors and regulatory elements in the geno...

Generation and characterization of a specific single-chain antibody against DSPP as a prostate cancer biomarker: Involvement of bioinformatics-based design of novel epitopes.

Isolation of specific single chain antibodies (scFvs) against key epitopes of cancer markers are applied for cancer immunotherapy and diagnosis. In this study following the prediction of the 3D structure of the DSP part of Dentin sialophosphoprotein (DSPP), the epitope was chosen using in silico programs. Panning process was applied to isolate specific human scFv against the epitope. PCR and DNA f...

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