Our official English website, www.x-mol.net, welcomes your
feedback! (Note: you will need to create a separate account there.)
Microencapsulated Immunoassays for Detection of Cytokines in Human Blood.
ACS Sensors ( IF 8.2 ) Pub Date : 2019-02-18 , DOI: 10.1021/acssensors.8b01033
Ali Rahimian 1 , Christian Siltanen 2 , Hamid Feyzizarnagh 1 , Patricio Escalante 3 , Alexander Revzin 1
ACS Sensors ( IF 8.2 ) Pub Date : 2019-02-18 , DOI: 10.1021/acssensors.8b01033
Ali Rahimian 1 , Christian Siltanen 2 , Hamid Feyzizarnagh 1 , Patricio Escalante 3 , Alexander Revzin 1
Affiliation
![]() |
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 hydrogel shell was composed of poly(ethylene glycol) forming a thin (∼10 μm) immunoisolation layer protecting antibody-modified microbeads inside the capsule from immune cells on the outside. The microbeads were functionalized with antibodies against cytokines of interest: interferon (IFN)-γ and tumor necrosis factor (TNF)-α. While nonfouling, a hydrogel shell was permeable to cytokine molecules; these molecules were captured on microbeads and were detected with fluorescently labeled secondary antibodies. Calibration of encapsulated immunoassays with known concentrations of cytokines revealed a limit of detection of 14.8 and 14.4 pM for IFN-γ and TNF-α, respectively. We also demonstrated the concept of multi-cytokine detection by fabricating distinct populations of capsules carrying either anti-IFN-γ or anti-TNF-α microbeads and dispensing these capsules into a solution containing both cytokine types. Importantly, when placed into whole blood for 16 h, microcapsules were free of leukocytes, effectively protecting sensing beads from the blood components. To further demonstrate utility of this strategy, encapsulated microbeads were used for detection of IFN-γ in blood of patients with latent tuberculosis infection (LTBI) and unexposed healthy controls. When compared to gold standard technology (interferon gamma release assay or IGRA), our encapsulated immunoassay accurately predicted LTBI diagnosis in 11 out of 14 patients. Overall, encapsulation of immunoassays represents a promising strategy for keeping sensing elements operational in a highly fouling complex environment such as blood.
中文翻译:
用于检测人血中细胞因子的微囊免疫测定。
细胞因子是由血液中的白细胞产生的,可用作恶性肿瘤或感染的指标。这项研究的目的是开发一种在未加工的全人类血液中免疫传感细胞因子的策略。微流体液滴的产生被用来制造直径约400μm的微胶囊,该微胶囊具有水凝胶壳和包含感测微珠的水核心。水凝胶壳由聚乙二醇组成,形成薄的免疫隔离层(约10μm),可保护胶囊内抗体修饰的微珠免受外部免疫细胞的侵害。用针对目标细胞因子的抗体功能化微珠:干扰素(IFN)-γ和肿瘤坏死因子(TNF)-α。在不结垢的同时,水凝胶壳对细胞因子分子具有渗透性。这些分子被捕获在微珠上,并用荧光标记的二抗检测。用已知浓度的细胞因子对包封的免疫测定进行校准,结果表明,IFN-γ和TNF-α的检出限分别为14.8和14.4 pM。我们还通过制造携带抗IFN-γ或抗TNF-α微珠的不同胶囊种群并将这些胶囊分配到包含两种细胞因子类型的溶液中,证明了多细胞因子检测的概念。重要的是,当将其置于全血中16 h时,微胶囊中没有白细胞,有效地保护了感测珠不受血液成分的影响。为了进一步证明此策略的实用性,包囊的微珠用于检测潜伏性结核感染(LTBI)和未暴露的健康对照者血液中的IFN-γ。与金标准技术(干扰素γ释放测定法或IGRA)相比,我们的封装免疫测定法可准确预测14例患者中11例的LTBI诊断。总体而言,免疫测定的封装代表了一种有前途的策略,可以使传感元件在血液等高度污染的复杂环境中保持正常运行。
更新日期:2019-02-18
中文翻译:
![](https://scdn.x-mol.com/jcss/images/paperTranslation.png)
用于检测人血中细胞因子的微囊免疫测定。
细胞因子是由血液中的白细胞产生的,可用作恶性肿瘤或感染的指标。这项研究的目的是开发一种在未加工的全人类血液中免疫传感细胞因子的策略。微流体液滴的产生被用来制造直径约400μm的微胶囊,该微胶囊具有水凝胶壳和包含感测微珠的水核心。水凝胶壳由聚乙二醇组成,形成薄的免疫隔离层(约10μm),可保护胶囊内抗体修饰的微珠免受外部免疫细胞的侵害。用针对目标细胞因子的抗体功能化微珠:干扰素(IFN)-γ和肿瘤坏死因子(TNF)-α。在不结垢的同时,水凝胶壳对细胞因子分子具有渗透性。这些分子被捕获在微珠上,并用荧光标记的二抗检测。用已知浓度的细胞因子对包封的免疫测定进行校准,结果表明,IFN-γ和TNF-α的检出限分别为14.8和14.4 pM。我们还通过制造携带抗IFN-γ或抗TNF-α微珠的不同胶囊种群并将这些胶囊分配到包含两种细胞因子类型的溶液中,证明了多细胞因子检测的概念。重要的是,当将其置于全血中16 h时,微胶囊中没有白细胞,有效地保护了感测珠不受血液成分的影响。为了进一步证明此策略的实用性,包囊的微珠用于检测潜伏性结核感染(LTBI)和未暴露的健康对照者血液中的IFN-γ。与金标准技术(干扰素γ释放测定法或IGRA)相比,我们的封装免疫测定法可准确预测14例患者中11例的LTBI诊断。总体而言,免疫测定的封装代表了一种有前途的策略,可以使传感元件在血液等高度污染的复杂环境中保持正常运行。