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Lymphoma cell isolation using multifunctional magnetic nanoparticles: antibody conjugation and characterization

机译：使用多功能磁性纳米颗粒分离淋巴瘤细胞：抗体偶联和表征

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The early detection of B-cell lymphoma and non-Hodgkin's lymphoma has a wide impact on the diagnosis and therapy of lymphoma patients. Capturing and sorting tumour cells with magnetic nanoparticles (MNPs) has received considerable attention in recent years. Despite these successes, the efficient isolation of circulating tumour cells from complex biological fluids is still under development for the early diagnosis of lymphoma. In this study, MNPs are functionalized with anti-CD20 antibodies using an avidin-biotin linkage, with the aim of achieving specific cancer cell detection and efficient isolation. Anti-CD20 antibody-conjugated MNPs (Ab MNPs) could specifically target CD20-expressing lymphoma cells. The capture efficiency of the Ab MNPs in the lymphoma cell line was >95% with regard to the mixture of two cell lines, as confirmed by flow cytometry and confocal microscopy. Transmission electron microscopy confirmed that the conjugation of an antibody with the MNPs increased the size from 12 to 47 nm. The surface charge of the Ab MNPs was examined by using zeta potential measurements. Furthermore, Prussian blue staining was performed to confirm the interaction of Ab MNPs with the targeted lymphoma cells. Our results indicated that the receptor recognition ability of the antibody was fully retained after conjugation with MNPs. In conclusion, anti-CD20 MNPs can be used for very sensitive detection and quick isolation of CD20-positive lymphoma cells among mixed cells by using only a permanent magnet.
机译：B细胞淋巴瘤和非霍奇金淋巴瘤的早期发现对淋巴瘤患者的诊断和治疗具有广泛的影响。近年来，利用磁性纳米颗粒（MNP）捕获和分选肿瘤细胞已受到相当大的关注。尽管取得了这些成功，但仍在开发从复杂生物液中有效分离循环肿瘤细胞以早期诊断淋巴瘤的方法。在这项研究中，使用抗生物素蛋白-生物素连接将MNPs与抗CD20抗体功能化，目的是实现特异性癌细胞检测和有效分离。抗CD20抗体偶联的MNP（Ab MNP）可以特异性地靶向表达CD20的淋巴瘤细胞。流式细胞术和共聚焦显微镜证实，相对于两种细胞系的混合物，Ab MNPs在淋巴瘤细胞系中的捕获效率> 95％。透射电子显微镜证实，抗体与MNP的缀合将尺寸从12nm增加至47nm。通过使用ζ电位测量来检查Ab MNP的表面电荷。此外，进行普鲁士蓝染色以确认Ab MNP与靶向淋巴瘤细胞的相互作用。我们的结果表明，与MNP结合后，抗体的受体识别能力被完全保留。总之，仅使用永磁体，抗CD20 MNP可用于非常灵敏的检测和快速分离混合细胞中CD20阳性淋巴瘤细胞。

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