Cytokine assays: An assessment of the preparation and treatment of blood and tissue samples

Keustermans, Genoveva; Hoeks, Sanne B.E.A.; Meerding, Jenny; Prakken, Berent J.; Jager, Wilco de

doi:10.1016/j.ymeth.2013.04.005

Cited by 139 publications

(106 citation statements)

References 59 publications

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“…Sample preparation and treatment (sample collection technique, storage duration, freeze-thaw exposure, etc) dramatically affect assay performance (see recent review by Keustermans et al (Keustermans et al, 2013) for an excellent comprehensive review of this topic). As our samples were collected in a standardized manner and fresh, newly frozen aliquots of plasma or BAL were used for the study, we did not test sample handling variations as part of this study.…”

Section: Discussionmentioning

confidence: 99%

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Bastarache

Koyama

Wickersham

et al. 2014

Journal of Immunological Methods

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Despite the widespread use of multiplex immunoassays, there are very few scientific reports that test the accuracy and reliability of a platform prior to publication of experimental data. Our laboratory has previously demonstrated the need for new assay platform validation prior to use of biologic samples from large studies in order to optimize sample handling and assay performance. In this study, our goal was to test the accuracy and reproducibility of an electrochemiluminescent multiplex immunoassay platform (Meso Scale Discovery, MSD®) and compare this platform to validated, singleplex immunoassays (R&D Systems®) using actual study subject (human plasma and mouse bronchoalveolar lavage fluid (BALF) and plasma) samples. We found that the MSD platform performed well on intra- and inter-assay comparisons, spike and recovery and cross-platform comparisons. The mean intra-assay CV% and range for MSD was 3.49 (0.0-10.4) for IL-6 and 2.04 (0.1-7.9) for IL-8. The correlation between values for identical samples measured on both MSD and R&D was R=0.97 for both analytes. The mouse MSD assay had a broader range of CV% with means ranging from 9.5-28.5 depending on the analyte. The range of mean CV% was similar for single plex ELISAs at 4.3-23.7 depending on the analyte. Regardless of species or sample type, CV% was more variable at lower protein concentrations. In conclusion, we validated a multiplex electrochemiluminscent assay system and found that it has superior test characteristics in human plasma compared to mouse BALF and plasma. Both human and MSD assays compared favorably to well-validated singleplex ELISA's

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Section: Discussionmentioning

confidence: 99%

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Bastarache

Koyama

Wickersham

et al. 2014

Journal of Immunological Methods

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“…Tetraspanins are organized into membrane microdomains which appear to play an important role in exosome biogenesis [21]. The ESCRT complex represents the machinery for exosome sorting, packaging and transport [23]. It consists of four membrane-bound aggregates of ubiquitinated proteins [15].…”

Section: Exosome Biogenesis In Cancermentioning

confidence: 99%

“…Newer data suggest that the profiles of the exosome cargo may differ substantially from those in parent cells. In one study, the relative quantities of proteins and nucleic acids in exosomes were found to be different from those in the parent cells [23]. Such differences may reflect the existence of a sorting process taking place during packaging of the cellular content into exosomes, although the exact way of how this process is regulated is still unknown.…”

Section: Exosome Biogenesis In Cancermentioning

confidence: 99%

Tumor-Derived Exosomes and Their Role in Cancer Progression

Whiteside

2016

Advances in Clinical Chemistry

621

492

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Tumor cells actively produce, release and utilize exosomes to promote tumor growth. Mechanisms through which tumor-derived exosomes subserve the tumor are under intense investigation. These exosomes are information carriers, conveying molecular and genetic messages from tumor cells to normal or other abnormal cells residing at close or distant sites. Tumor-derived exosomes are found in all body fluids. Upon the contact with target cells, they alter phenotypic and functional attributes of recipients, reprogramming them into active contributors to angiogenesis, thrombosis, metastasis and immunosuppression. Exosomes produced by tumors carry cargos that in part mimic contents of parent cells and are of potential interest as non-invasive biomarkers of cancer. Their role in inhibiting the host antitumor responses and in mediating drug resistance is important for cancer therapy. Tumor-derived exosomes may interfere with cancer immunotherapy, but they also could serve as adjuvants and antigenic components of antitumor vaccines. Their biological roles in cancer development or progression as well as cancer therapy suggest that tumor-derived exosomes are critical components of oncogenic transformation.

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“…When considering cytokine measurement, several issues should be considered in order to maintain cytokine stability: 1) samples should be stored at low temperatures (f -80uC); 2) multiple freezethaw cycles should be avoided; and 3) while most cytokines remain stable within a 2-year period, longer term storage may alter cytokine levels as a result of cross-reactivity between protein epitopes [11]. Despite the fact that samples were kept at -180uC and they did not undergo any freeze-thaw cycles, as used samples do not re-enter the biobank, the storage time was long (5-8 years).…”

mentioning

confidence: 99%

Proinflammatory cytokine levels are linked to death in pulmonary arterial hypertension

Chabot²,

et al. 2013

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Cytokine assays: An assessment of the preparation and treatment of blood and tissue samples

Cited by 139 publications

References 59 publications

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Validation of a multiplex electrochemiluminescent immunoassay platform in human and mouse samples

Tumor-Derived Exosomes and Their Role in Cancer Progression

Proinflammatory cytokine levels are linked to death in pulmonary arterial hypertension

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