从纳米免疫疗法的角度看卵清蛋白中的内毒素污染,WIREs Nanomedicine and Nanobiotechnology

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从纳米免疫疗法的角度看卵清蛋白中的内毒素污染
WIREs Nanomedicine and Nanobiotechnology ( IF 6.9 ) Pub Date : 2021-08-10 , DOI: 10.1002/wnan.1747
Guibin Pang ₁ , Yun Liu ₂ , Yangyun Wang ₃ , Yong Wang ₃ , Fujun Wang ₂ , Jian Zhao ₁ , Leshuai W. Zhang ₁

Affiliation

卵清蛋白 (OVA) 是一种模型抗原，通常掺入巧妙设计的纳米粒子中，用于递送到抗原呈递细胞 (APC) 中，旨在研究含有免疫调节化合物的纳米粒子的免疫活性和治疗功效。然而，在纳米免疫疗法中观察到的免疫反应可能出乎意料地来自纳米颗粒中OVA的内毒素杂质。文献回顾表明，大多数研究人员没有注意到无内毒素 OVA 在用于纳米免疫疗法研究时的重要性。来自 Sigma-Aldrich 的低至 5 μg/ml OVA 的浓度（包含 0.625 ng/ml 内毒素）能够激活 APC，例如树突状细胞和巨噬细胞。在这里，我们建议 OVA 或成品纳米产品中的内毒素杂质应通过鲎变形细胞裂解物 (LAL) 和基于细胞的测定来确定，以确保纳米粒子的无内毒素质量。OVA中的内毒素可以通过内毒素去除柱和相分离的方法去除，可以购买不含内毒素的OVA。这一观点提醒研究人员注意 OVA 的内毒素杂质可能会转移到成品纳米颗粒中，并引入不利的免疫调节功能，并导致假阳性结果。内毒素水平最低的 OVA 应用于纳米免疫疗法研究，以准确反映纳米颗粒对免疫系统的真实影响。这一观点提醒研究人员注意 OVA 的内毒素杂质可能会转移到成品纳米颗粒中，并引入不利的免疫调节功能，并导致假阳性结果。内毒素水平最低的 OVA 应用于纳米免疫疗法研究，以准确反映纳米颗粒对免疫系统的真实影响。这一观点提醒研究人员注意 OVA 的内毒素杂质可能会转移到成品纳米颗粒中，并引入不利的免疫调节功能，并导致假阳性结果。内毒素水平最低的 OVA 应用于纳米免疫疗法研究，以准确反映纳米颗粒对免疫系统的真实影响。

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Endotoxin contamination in ovalbumin as viewed from a nano-immunotherapy perspective

Ovalbumin (OVA) is a model antigen commonly incorporated in smartly designed nanoparticles for delivery into antigen-presenting cells (APC), aiming to investigate the immune activity and therapeutic efficacy of nanoparticles that contain immunoregulatory compounds. However, the immunoresponse observed in nano-immunotherapy may unexpectedly arise from endotoxin impurity of OVA in the nanoparticles. Literature review shows that most researchers did not notice the importance of endotoxin-free OVA when used in nano-immunotherapy studies. Concentration at as low as 5 μg/ml OVA from Sigma-Aldrich (contains 0.625 ng/ml endotoxin) was able to activate APC such as dendritic cells and macrophages. Here, we proposed that the endotoxin impurity in OVA or the finished nanoproducts should be determined by both Limulus Amebocyte Lysate (LAL) and cell-based assay, to ensure the endotoxin-free quality of the nanoparticles. The endotoxin in OVA can be removed by endotoxin removal column and phase separation methods and endotoxin-free OVA can be purchased. This perspective alerts the researchers of endotoxin impurity of OVA that may transfer into the finished nanoparticles and introduce an unfavorable immunoregulatory function with false-positive results. OVA with minimal endotoxin level should be used in nano-immunotherapy studies to accurately reflect the true effects of nanoparticles on the immune system.

更新日期：2021-08-10

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