Nanocrystal quantum dot-conjugated anti-myeloperoxidase antibody as the detector of activated neutrophils
Nanocrystal quantum dot-conjugated anti-myeloperoxidase antibody as the detector of activated neutrophils
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DOI：
10.1109/tnb.2007.909008
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发表时间：
2007-12-01
影响因子：
3.9
通讯作者：
Yamamoto, Kenji
中科院分区：
文献类型：
--
作者：
Hoshino, Akiyoshi;Nagao, Tomokazu;Yamamoto, Kenji
荧光双量子点（QD）通过利用其荧光特性已被应用于广泛的生物学研究。在这里，我们表明量子点结合抗体对中性粒细胞过氧化物酶，髓过氧化物酶（MPO）。我们设计了一种新的方法将量子点偶联到抗体上，而不损失任何抗体功能，包括它们的抗原识别和Fc受体结合活性。当我们将抗MPO抗体（Ab）与常规有机探针一起应用于活细胞的免疫染色时，由于MPO酶活性产生活性氧，抗体失去荧光。我们的量子点结合的抗MPO（α-MPO-量子点）可以检测激活的中性粒细胞表面的MPO。此外，抗-MPO-QD不与灭活的中性粒细胞反应。总之，我们证明，抗体可视化的MPO在中性粒细胞表面的表达后，刺激与促炎细胞因子。总而言之，这些技术有可能通过免疫染色和流式细胞术分析揭示中性粒细胞的活化，作为体外诊断中性粒细胞活化的有力工具。
Fluorescent nanocrystal quantum dots (QDs) have been applied to a wide range of biological studies by taking advantage of their fluorescence properties. Here we show that QDs conjugated with antibody against neutrophil peroxidase, myeloperoxidase (MPO). We designed a novel method to conjugate QDs to antibody without losing any antibody function including their antigen recognizing and Fc-receptor binding activities. When we applied anti-MPO antibody (Ab) with conventional organic probes in the case of immunostaining of living cells, the antibodies lost their fluorescence because of MPO enzymic activity to produce reactive oxygen species. Our QD-conjugated anti-MPO (alpha-MPO-QDs) can detect MPO on the surface of activated neutrophils. In addition, anti-MPO-QDs did not react to the inactivated neutrophils. In conclusion, we demonstrated that antibody visualized the expression of MPO on the neutrophil surface after stimulation with proinflammatory cytokines. Taken together, these techniques have the possibility that QDs can reveal the activation of neutrophils by immunostaining and flow cytometric analysis as a powerful tool for diagnosis of the neutrophil activation in vitro.