ELISA-based detection of immunoglobulins against extracellular vesicles in blood plasma

Tom A. P. Driedonks, Linglei Jiang, Olesia Gololobova, Zhaohao Liao, Kenneth W. Witwer
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Abstract

Extracellular vesicles (EVs) are intensively investigated for their therapeutic potential and application as drug delivery vehicle. A broad perception of favourable safety profiles and low immunogenicity make EVs an attractive alternative to synthetic nanoparticles. We recently showed that repeated intravenous administration of human cell-derived EVs into pig-tailed macaques unexpectedly elicited antibody responses after three or more injections. This coincided with decreasing EV circulation time, and may thus hamper successful EV-mediated cargo delivery into tissues. Here, we share the custom ELISA protocol that we used to measure such antibody responses. This protocol may help other researchers evaluate immune responses to EV-based therapies in preclinical studies.

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基于 ELISA 法检测血浆中针对细胞外囊泡的免疫球蛋白
细胞外囊泡(EVs)因其治疗潜力和作为药物输送载体的应用而受到广泛关注。人们普遍认为细胞外囊泡具有良好的安全性和低免疫原性,因此是合成纳米颗粒的一种有吸引力的替代品。我们最近的研究表明,向猪尾猕猴反复静脉注射人体细胞衍生的 EVs,在注射三次或三次以上后,会意外地引起抗体反应。这与EV循环时间的缩短不谋而合,因此可能会阻碍EV介导的货物成功输送到组织中。在此,我们与大家分享用于测量此类抗体反应的定制 ELISA 方案。该方案可帮助其他研究人员在临床前研究中评估基于 EV 的疗法的免疫反应。
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