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刘辉荣, 徐宏, 古宏晨, 卢瑛, 王玲, 侯盛. 简便高效分离细胞新型免疫磁珠制备[J]. 中国公共卫生, 2008, 24(11): 1349-1351. DOI: 10.11847/zgggws2008-24-11-38
引用本文: 刘辉荣, 徐宏, 古宏晨, 卢瑛, 王玲, 侯盛. 简便高效分离细胞新型免疫磁珠制备[J]. 中国公共卫生, 2008, 24(11): 1349-1351. DOI: 10.11847/zgggws2008-24-11-38
LIU Hui-rong, XU Hong, GU Hong-chen, . New immunomagnetic beads used for cell separation simply and high efficiently[J]. Chinese Journal of Public Health, 2008, 24(11): 1349-1351. DOI: 10.11847/zgggws2008-24-11-38
Citation: LIU Hui-rong, XU Hong, GU Hong-chen, . New immunomagnetic beads used for cell separation simply and high efficiently[J]. Chinese Journal of Public Health, 2008, 24(11): 1349-1351. DOI: 10.11847/zgggws2008-24-11-38

简便高效分离细胞新型免疫磁珠制备

New immunomagnetic beads used for cell separation simply and high efficiently

  • 摘要: 目的 采用碳二亚胺/N-羟基琥珀酰亚胺(EDC/NHS)方法在羧基磁珠表面偶联抗体,制备可高效分离细胞的免疫磁珠。方法 用EDC和NHS活化磁珠表面羧基,活化的羧基再与抗体上氨基进行反应,从而将抗体偶联于磁珠表面,获得免疫磁珠。使用高性能纳米粒度分析仪(HPPS)、二辛可酸(BCA)蛋白定量试剂盒、流式细胞仪、透射电镜(TEM)表征磁珠的粒径、磁珠表面联接的抗体量及其免疫活性。结果 HPPS检测磁珠平均水力学粒径为110nm;磁珠表面偶联52.4μg抗CD11a+抗体/mg磁珠。经磁分离后细胞的流式分析结果表明,CD11a+免疫磁珠可以有效分离髓系白血病细胞系(KG-1a)细胞,免疫磁珠均匀结合于细胞表面,且不影响细胞的活性。结论 成功制备可用于细胞分离且不影响分离后细胞活性的新型免疫磁珠。

     

    Abstract: Objective To prepare the immunomagnetic bead used for cell separation simply and high efficiently.Methods The immunomagnetic beads were prepared by the reaction between the amidos of the antibodies and the carboxyl on the surface of magnetic beads whose carboxyl groups were activated by EDC/NHS.The properties of magnetic beads were studied by dynamic light scattering size analyse,BCA protein quantification assay,flow cytometer and TEM.Results The diameter of the magnetic beads was 110 nm.There were 5214μg CD11a+antibodies lebeled on the per mg magnetic beads.FACS results indicated that KG21a cell were separated efficiently by the immunomagnetic beads.TEM results indicated that the immunomagnetic beads were combined on the surface of the cell.The cultivation and microscopic observation indicated that the immunomagnetic beads didn't affect the activation of the cell.Conclusion The new kind immunomagnetic beads which could be used for cell separation without negative effect on cell's activation is prepared successfully.

     

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