ELISAs and similar immunoassays are a backbone of biomedical research and clinical practice. Here we review the major factors to consider in the development and application of ultrasensitive ELISAs for analysis of human immune responses in plasma, serum, urine, or tissue culture supernatants. We focus on cytokine and chemokine biomarkers of health and chronic inflammatory diseases including allergy, asthma, autoimmunity, and cardiovascular disease. Detailed protocols for ELISA and Meso Scale Discovery assays (an improved variant of ELISA) are provided for 15 cytokines and 11 chemokines that play immune-regulatory roles in human innate and adaptive immunity. Protocols have been individually optimized to yield ultrasensitive limits of detection and quantification. Major factors enhancing immunoassay sensitivity, precision, and reproducibility, as well as key pitfalls in assay design and execution, are critically reviewed.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-4939-9591-2_7 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Efficacy and Possible Mechanism(s) of Action of Gallium Tetraphenylporphyrin Nanoparticles against HIV-TB Coinfection in an Granuloma Structure Model.
ACS Infect Dis
December 2024
Department of Pathology, Microbiology and Immunology, College of Medicine, University of Nebraska Medical Center, Omaha, Nebraska 68198, United States.
Coinfection of (Mtb) and human immunodeficiency virus-1 (HIV) is a significant public health concern. Treatment is challenging due to prolonged duration of therapy and drug interactions between antiretroviral therapy (ART) and anti-TB drugs. Noniron gallium -tetraphenyl porphyrin (GaTP), a heme mimetic, has shown broad antimicrobial activity.
View Article and Find Full Text PDFChidan Tuihuang granule modulates gut microbiota to influence NOD1/RIPK2 pathway in cholestatic liver injury recovery.
Phytomedicine
December 2024
Department of Pharmaceutical Biology, Institute of Pharmaceutical and Biomedical Sciences, Johannes Gutenberg University, Mainz, Germany. Electronic address:
Cholestatic liver injury (CLI), which occurs if bile acids are imbalanced and the liver becomes inflamed, is difficult to treat effectively OBJECTIVE: We investigated how the Chinese patent medicine Chidan Tuihuang granule (CDTH) ameliorates cholestatic liver injury with a focus on its effects on the NOD1/RIPK2 pathway and intestinal flora METHODS: We used an ANIT-induced SD rat model of CLI to evaluate the therapeutic effects of CDTH. The experimental design included control, model, UDCA (ursodeoxycholic acid) and CDTH treatment groups. UHPLC-Q-Orbitrap-HRMS was used to analyse the blood components of CDTH.
View Article and Find Full Text PDFSystemic increase in IL-26 is associated with severe COVID-19 and comorbid obstructive lung disease.
Front Immunol
October 2024
Division of Lung and Airway Research, Institute of Environmental Medicine, and the Center for Molecular Medicine, Karolinska Institutet, Stockholm, Sweden.
Background: IL-26 is a key mediator of pulmonary host defense given its abundant expression in human airways and its established antibacterial properties. Moreover, recent studies indicate that IL-26 can also inhibit viral replication. Along these lines, we have previously reported an increase in the plasma concentration of IL-26 among patients with acute COVID-19 that is linked to harmful hyperinflammation.
View Article and Find Full Text PDFThe Interruption of Transmission of Onchocerciasis in Abia, Anambra, Enugu, and Imo States, Nigeria: The Largest Global Onchocerciasis Stop-Treatment Decision to Date.
Pathogens
August 2024
The Carter Center, Atlanta, GA 30307, USA.
Onchocerciasis causes severe morbidity in sub-Saharan Africa. Abia, Anambra, Enugu, and Imo states of Nigeria were historically classified meso- or hyperendemic and eligible for ivermectin mass drug administration (MDA). After ≥25 years of annual and biannual MDA, serological and entomological assessments were conducted to determine if transmission was interrupted.
View Article and Find Full Text PDFA Data Driven Strategy and Case Study for Implementation of Singlicate Analysis in Ligand Binding Assays Used for PK Quantitation.
AAPS J
July 2024
Clinical Bioanalytics, Pfizer Inc, Andover, USA.
Duplicate analysis has been a conventional practice in the industry for ligand-binding assays (LBA), particularly for plate-based platforms like Enzyme-linked immunosorbent assay (ELISA) and Meso Scale Discovery (MSD) assays. Recent whitepapers and guidance have opened a door to exploring the implementation of single-well (singlicate) analysis approach for LBAs. Although the bioanalytical industry has actively investigated the suitability of singlicate analysis, applications in supporting regulated LBA bioanalysis are limited.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!