Filatov M V, Landa S B, Pantina R A, Garmaĭ Iu P
Klin Lab Diagn. 2010 Dec(12):35-43.
Laser correlation spectroscopy, atomic force microscopy, and immunoaffinity chromatography were used to characterize exosomes produced by different human cells. The exosomes secreted into a culture medium by normal fibroblasts, dendritic cells, lymphocytes, as well as malignant cells obtained from tumors of various tissue origins. The similar investigations were made for exosomes detectable in plasma and cerebrospinal fluid. The dynamic light scattering technique has demonstrated that the exosomes from different sources are homogenous and similar in size of the order of 20 and 90 nm. The exceptional homogeneity of exosomes was confirmed by atomic force microscopy. The immunoaffinity method has shown that all the exosomes under study carry antigenic determinants recognizable by antibodies to the major histocompatibility complex of type 1 (HLA-ABC). A method is proposed for evidence-based detection of exosomes in various biological fluids. For this, dynamic light scattering detects 20- and 90-nm particles and whether they can be removed by immunoaffinity chromatography with HLA-ABC antibodies is checked.
激光相关光谱法、原子力显微镜和免疫亲和色谱法被用于表征不同人类细胞产生的外泌体。外泌体由正常成纤维细胞、树突状细胞、淋巴细胞以及从各种组织来源的肿瘤中获得的恶性细胞分泌到培养基中。对血浆和脑脊液中可检测到的外泌体进行了类似的研究。动态光散射技术表明,来自不同来源的外泌体是均匀的,大小在20到90纳米左右相似。原子力显微镜证实了外泌体具有非凡的均匀性。免疫亲和方法表明,所有研究的外泌体都携带可被针对1型主要组织相容性复合体(HLA-ABC)的抗体识别的抗原决定簇。提出了一种基于证据的检测各种生物流体中外泌体的方法。为此,动态光散射检测20纳米和90纳米的颗粒,并检查它们是否可以通过用HLA-ABC抗体的免疫亲和色谱法去除。