Article

Lateral flow assay for simultaneous detection of cellular- and humoral immune responses

Department of Molecular Cell Biology Leiden University Medical Center, Einthovenweg 20, 2333 ZC Leiden, The Netherlands.
Clinical Biochemistry (Impact Factor: 2.23). 07/2011; 44(14-15):1241-6. DOI: 10.1016/j.clinbiochem.2011.06.983
Source: PubMed

ABSTRACT The development of a cytokine detection assay suitable for detection of multiple biomarkers for improved diagnosis of mycobacterial diseases.
A lateral flow (LF) assay to detect IL-10 was developed utilizing the up-converting phosphor (UCP) reporter-technology. The assay was evaluated using blood samples of leprosy patients. Multiplex applications were explored targeting: 1) IL-10 and IFN-γ in assay buffer; 2) IL-10 and anti-phenolic glycolipid (PGL-I) antibodies in serum from leprosy patients.
Detection of IL-10 below the targeted level of 100pg/mL in serum was shown. Comparison with ELISA showed a quantitative correlation with R(2) value of 0.92. Multiplexing of cytokines and simultaneous detection of cytokine and antibody was demonstrated.
The UCP-LF IL-10 assay is a user-friendly, rapid alternative for IL-10 ELISAs, suitable for multiplex detection of different cytokines and can be merged with antibody-detection assays to simultaneously detect cellular- and humoral immunity.

Download full-text

Full-text

Available from: Paul Corstjens, Jul 19, 2015
2 Followers
 · 
281 Views
  • Source
    • "The combination of the UCP reporter technology with the user-friendly and rapid LF-based immunochromatography has shown better sensitivity than ELISA-based assays, and several UCP-LF applications to detect different types of biomolecules have already been described (references recapped in Corstjens et al. 2011 [18]). In the current study, the focus is on IFX and IBD, but the results indicate that the assay in its current form is directly applicable to other biologicals (immunotherapeutics) and diseases as e.g. "
    [Show abstract] [Hide abstract]
    ABSTRACT: Monitoring levels of biologicals against tumor necrosis factor (TNF) has been suggested to improve therapeutic outcomes in inflammatory bowel diseases (IBDs). This pilot study describes a rapid lateral flow (LF)-based assay for on-site monitoring of serum trough levels of humanized monoclonal antibody infliximab (IFX). The applied chromatographic method utilizes sequential flows of diluted serum, wash buffer, and an immunoglobulin generic label on LF strips with a Test line comprised of TNF-α. The successive flows permitted enrichment of IFX at the Test line before the label was applied. The label, luminescent upconverting phosphor (UCP) particles coated with protein-A, emits a 550-nm visible light upon excitation with 980-nm infrared light. IFX concentrations were determined through measurement of UCP fluorescence at the Test line. The assay was optimized to detect IFX levels as low as 0.17 μg/mL in serum. For patients with IBD, this limit is appropriate to detect levels associated with loss of response (0.5μg IFX/mL). The assay was evaluated with clinical samples from patients with Crohn’s disease and correlated well within the physiologically relevant range from 0.17 to 10 μg/mL with an IFX-specific ELISA. Performance of the assay was further successfully validated with samples from blood donors, IFX negative IBD patients, and rheumatoid arthritis patients that had developed anti-IFX antibodies. Because of its generic nature, the assay is suited for detecting most therapeutic anti-TNF-α monoclonal antibodies.
    Analytical and Bioanalytical Chemistry 07/2013; DOI:10.1007/s00216-013-7154-0 · 3.58 Impact Factor
  • Source
    Leprosy review 12/2011; 82(4):340-3. · 0.59 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Lateral flow (LF) tests are frequently used for rapid delivery of qualitative diagnostic results. The detection sensitivity of these immunochromatography assays is largely dependent on the reporter technology. Many LF assays utilize reporters suitable for a visual interpretation of the test, implying less-sensitive detection than fluorescent reporter-based technologies that require LF strip readers to determine the test result. Here we describe the implementation of an unconventional and ultrasensitive fluorescent reporter technology: upconverting phosphor reporter particles. The reporter particles are excited with low energy 980 nm near infrared light and emit higher energy visible light that, similar to other fluorescent labels, allows a quantitative analysis of the test result. Detailed procedures regarding the LF strip production, bioconjugation of the reporter particles and the actual LF assay are described. The result of the UCP-LF assay is determined with dedicated readers and software, including a lightweight portable reader.
    Methods in cell biology 01/2012; 112:203-234. DOI:10.1016/B978-0-12-405914-6.00011-1 · 1.44 Impact Factor
Show more