Improved Methods for Quantifying Human Chemokine and Cytokine Biomarker Responses: Ultrasensitive ELISA and Meso Scale Electrochemiluminescence Assays

Stefura, William P.; Graham, Caroline; Lotoski, Larisa; HayGlass, Kent T.

doi:10.1007/978-1-4939-9591-2_7

Cited by 13 publications

(12 citation statements)

References 13 publications

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“…Biomarkers include salivary cortisol, a measure of hypothalamic-pituitary-adrenal axis activity, collected at wake, 30 min post-wake, and bedtime for 3 consecutive days according to well-established procedures; 54 and systemic markers of inflammation, assessed by using a Meso Scale Discovery electrochemiluminescence assay 55 to determine 14 plasma markers of inflammation, including interleukin 1, interleukin 6, interleukin 17, tumor necrosis factor, and eotaxin concentrations. We also store serum, plasma, and buffy coat samples at each time point as well as DNA for future analyses.…”

Section: Assessmentsmentioning

confidence: 99%

Mindfulness, Education, and Exercise for age-related cognitive decline: Study protocol, pilot study results, and description of the baseline sample

et al. 2020

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Background/Aims Age-related cognitive decline is a pervasive problem in our aging population. To date, no pharmacological treatments to halt or reverse cognitive decline are available. Behavioral interventions, such as physical exercise and Mindfulness-Based Stress Reduction, may reduce or reverse cognitive decline, but rigorously designed randomized controlled trials are needed to test the efficacy of such interventions. Methods Here, we describe the design of the Mindfulness, Education, and Exercise study, an 18-month randomized controlled trial that will assess the effect of two interventions—mindfulness training plus moderate-to-vigorous intensity exercise or moderate-to-vigorous intensity exercise alone—compared with a health education control group on cognitive function in older adults. An extensive battery of biobehavioral assessments will be used to understand the mechanisms of cognitive remediation, by using structural and resting state functional magnetic resonance imaging, insulin sensitivity, inflammation, and metabolic and behavioral assessments. Results We provide the results from a preliminary study (n = 29) of non-randomized pilot participants who received both the exercise and Mindfulness-Based Stress Reduction interventions. We also provide details on the recruitment and baseline characteristics of the randomized controlled trial sample (n = 585). Conclusion When complete, the Mindfulness, Education, and Exercise study will inform the research community on the efficacy of these widely available interventions improve cognitive functioning in older adults.

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Section: Assessmentsmentioning

confidence: 99%

Mindfulness, Education, and Exercise for age-related cognitive decline: Study protocol, pilot study results, and description of the baseline sample

et al. 2020

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“…A pertinent detail is also at the level of efficacy of the detection antibodies in ELISA used to measure cytokine concentrations in plasma, in particular for very low cytokine concentrations such as those of type I IFN. 34 Hence, the standardization of the reagents used to set up the ELISAs and similar immunoassays (multiplex technologies, electrochemiluminescence, single-molecular array) using monoclonal antibodies [113][114][115] will be of great importance to allow efficient comparisons of data without forgetting the possible false-positive results in plasma/serum due to heterophilic antibodies (i.e., human anti-mouse antibodies) possibly present in human plasma/serum 116,117 . Regarding the cytokine secretome and the complex functions of cytokines (pleiotropy, redundancy, the duality of actions), it seems of significant importance to have the overall profile of cytokines involved in each patient's disease.…”

Section: Why When and How To Evaluate Cytokines?mentioning

confidence: 99%

The evaluation of cytokines to help establish diagnosis and guide treatment of autoinflammatory and autoimmune diseases

Nézondet

Poubelle

Pelletier

2020

Journal of Leukocyte Biology

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Our knowledge of the role of cytokines in pathologic conditions has increased considerably with the emergence of molecular and genetic studies, particularly in the case of autoinflammatory monogenic diseases. Many rare disorders, considered orphan until recently, are directly related to abnormal gene regulation, and the treatment with biologic agents (biologics) targeting cytokine receptors, intracellular signaling or specific cytokines improve the symptoms of an increasing number of chronic inflammatory diseases. As it is currently impossible to systematically conduct genetic studies for all patients with autoinflammatory and autoimmune diseases, the evaluation of cytokines can be seen as a simple, less time consuming, and less expensive alternative. This approach could be especially useful when the diagnosis of syndromes of diseases of unknown etiology remains problematic. The evaluation of cytokines could also help avoid the current trial-and-error approach, which has the disadvantages of exposing patients to ineffective drugs with possible unnecessary side effects and permanent organ damages. In this review, we discuss the various possibilities, as well as the limitations of evaluating the cytokine profiles of patients suffering from autoinflammatory and autoimmune diseases, with methods such as direct detection of cytokines in the plasma/serum or following ex vivo stimulation of PBMCs leading to the production of their cytokine secretome. The patients' secretome, combined with biomarkers ranging from genetic and epigenetic analyses to immunologic biomarkers, may help not only the diagnosis but also guide the choice of biologics for more efficient and rapid treatments.

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“…For example, many commercial double-antibody sandwich ELISAs have been developed for the diagnosis of human and animal diseases [4,5]. To develop this assay, the use of capture and reporter-labeled detection antigen-specific antibodies is essential and must be produced in the initial step [6,7]. While most sandwich ELISA kits and housed-methods employ conventional polyclonal and monoclonal antibodies as indispensable reagents, they present several drawbacks, including limited amounts, difficulty in permanent storage, and required use of a secondary antibody [8,9].…”

Section: Introductionmentioning

confidence: 99%

Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

Zhu

et al. 2020

Preprint

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Background Traditional sandwich enzyme-linked immunosorbent assay (ELISA) using polyclonal and monoclonal antibodies as reagents presents several drawbacks, including limited amounts, difficulty in permanent storage, and required use of a secondary antibody. Nanobodies can be easily expressed with different systems and fused with several tags in their tertiary structure by recombinant technology, thus offering an effective detection method for diagnostic purposes. Recently, the fenobody (ferritin-fused nanobody) and RANbody (nanobody-fused reporter) have been designed and derived from the nanobody for developing the diagnostic immunoassays. However, there was no report about developing the sandwich ELISA using the fenobody and RANbody as pairing reagents. Results A platform for developing a sandwich ELISA utilizing fenobody as the capture antibody and RANbody as the detection antibody was firstly designed in the study. Newcastle disease virus (NDV) was selected as the antigen, from which 13 NDV-specific nanobodies were screened from an immunized Bactrian camel. Then, 5 nanobodies were selected to produce fenobodies and RANbodies. The best pairing of fenobodies (NDV-fenobody-4, 800 ng/well) and RANbodies (NDV-RANbody-49, 1:10) was determined to develop the sandwich ELISA for detecting NDV. The detection limits of the assay were determined to be 2 2 of hemagglutination (HA) titers and 10 ng of purified NDV particles. Compared with two commercial assays, the developed assay shows higher sensitivity and specificity. Meanwhile, it exhibits 98.7% agreement with the HA test and can detect the reference NDV strains belonging to Class II but not Class I. Conclusions In the presented study, the 13 anti-NDV nanobodies binding the NDV particles were first produced. Then, for the first time, the sandwich ELISA to detect the NDV in the different samples has been developed using the fenobody and RANbody as reagents derived from the nanobodies. Considering the rapidly increasing generation of nanobodies, the platform can reduce the cost of production for the sandwich ELISA and be universally used to develop assays for detecting other antigens.

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Improved Methods for Quantifying Human Chemokine and Cytokine Biomarker Responses: Ultrasensitive ELISA and Meso Scale Electrochemiluminescence Assays

Cited by 13 publications

References 13 publications

Mindfulness, Education, and Exercise for age-related cognitive decline: Study protocol, pilot study results, and description of the baseline sample

Mindfulness, Education, and Exercise for age-related cognitive decline: Study protocol, pilot study results, and description of the baseline sample

The evaluation of cytokines to help establish diagnosis and guide treatment of autoinflammatory and autoimmune diseases

Fenobody and RANbody-based sandwich enzyme-linked immunosorbent assay to detect Newcastle disease virus

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