Preparation of PEG-grafted immunomagnetoliposomes entrapping citrate stabilized magnetite particles and their application in CD34+ cell sorting.
Preparation of PEG-grafted immunomagnetoliposomes entrapping citrate stabilized magnetite particles and their application in CD34+ cell sorting.
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包埋柠檬酸盐稳定磁铁矿颗粒的 PEG 接枝免疫磁脂质体的制备及其在 CD34 细胞分选中的应用。
DOI：
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发表时间：
2001
影响因子：
3.9
通讯作者：
M. A. D. Madariaga
中科院分区：
文献类型：
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作者：
Joan Carles Domingo;M. Mercadal;Jordi Petriz;M. A. D. Madariaga
免疫磁系统已用于阳性选择细胞部分的混合物使用适当的表面标记，结果令人满意，如造血CD34+细胞。本文报道了负载柠檬酸磁铁矿稳定颗粒的聚乙二醇（PEG）免疫脂质体作为免疫磁分离载体的发展。采用DRV方法将磁性铁磁流体包封到peg脂质体中。磁性脂质体的脂质体大小约为450 nm，铁/脂质摩尔比为1.52+/-0.26，并保留在MiniMACS系统产生的磁场中。通过偶联My10 mAb制备了100 mAb/囊泡的抗CD34免疫磁脂质体，并在培养和与CD34细胞（CHO或Jurkat）混合的CD34+ KG-1a细胞中特异性结合。在不同初始CD34+ KG-1a细胞百分比的KG-1a/CHO或KG-1a/Jurkat混合细胞中进行磁性细胞分选。对于10(6)个阳性细胞和100微米的免疫磁脂质体，捕获效率为0.85%，与CD34+细胞的起始百分比无关。当CD34+细胞的起始百分比减少时，最终纯度的降低，取决于所使用的CD34-细胞系，表明my10免疫磁脂质体的非特异性细胞结合程度至关重要，在方法学方面，如起始CD34+细胞的数量。分离的CD34+细胞保留了活力，估计回收率为45-50%。
Immunomagnetic systems have been used for positive selection of a cell fraction from a mixture using appropriate surface markers with satisfactory results, as haematopoietic CD34+ cells. This work reports on the development of poly(ethylene glycol)-grafted (PEG) immunoliposomes loaded with citrate-magnetite stabilized particles as the separation vehicles for immunomagnetic separations. The magnetic ferrofluid was encapsulated into PEG-liposomes by the DRV methodology. The magnetoliposomes had a liposomal size of approximately 450 nm and a Fe/lipid molar ratio of 1.52+/-0.26, and were retained in the magnetic field created by the MiniMACS system. Anti-CD34 immunomagnetoliposomes with 100 mAb/vesicle were prepared by coupling the My10 mAb and bound specifically for CD34+ KG-1a cells in culture and in mixtures with CD34-cells (CHO or Jurkat). The magnetic cell sorting was carried out in cell mixtures KG-1a/CHO or KG-1a/Jurkat with different initial% of CD34+ Kg-1a cells. For 10(6) positive cells and 100 microM of immunomagnetoliposomes, the capture efficiency was > 85% and independent of the starting percentage of CD34+ cells. The decrease of the final purity, when the starting percentage of CD34+ cells decreases and, dependent of the CD34- cell line used, point to the degree of non-specific cell binding of My10-immunomagnetoliposomes as being crucial, among of the methodological aspects as the number of starting CD34+ cells. The CD34+ cells isolated retained the viability with an estimated recovery of 45-50%.
影响因子：
3.4
作者：
BOGDANOV, AA;MARTIN, C;BRADY, TJ
通讯作者：
BRADY, TJ
影响因子：
3.4
作者：
Du, H;Chandaroy, P;Hui, SW
通讯作者：
Hui, SW