Iron Oxide Nanoparticle Assisted Purification and Mass Spectrometry Based Proteolytic Mapping of Intact CD4+T Cells from Human Blood
Iron Oxide Nanoparticle Assisted Purification and Mass Spectrometry Based Proteolytic Mapping of Intact CD4+T Cells from Human Blood
复制标题
氧化铁纳米颗粒辅助纯化和基于质谱的人血完整 CD4 T 细胞的蛋白水解图谱
DOI：
10.1080/10826060802589510
复制
发表时间：
2008
影响因子：
2.9
通讯作者：
M. Mandal
中科院分区：
文献类型：
--
作者：
S. Mandal;A. Ghosh;M. Mandal
摘要氧化铁纳米颗粒作为临床应用已经有多年的历史。我们已经开发了一种快速免疫亲和分离方法的CD 4 +T细胞的混合细胞群的人血液使用氧化铁纳米粒子。抗CD 4抗体经表面修饰后附着在氧化铁纳米颗粒上。将抗体标记的氧化铁纳米颗粒珠与人血液的混合细胞群简单地孵育，并使用外部磁场纯化CD 4 +T细胞。通过荧光显微镜和流式细胞术检查纯化水平。纯化后的CD 4 +T细胞用胰蛋白酶消化不同时间段，并通过MALDI-TOF质谱分析产物，无需进一步分离或纯化，以获得其蛋白质组图谱。数据库检索显示许多肽质量与T细胞肽质量特异性匹配。这些结果表明氧化铁纳米颗粒可用于CD 4 +T细胞纯化，并且基于质谱的蛋白水解指纹对于从整个细胞表面鉴定推定的表面生物标志物是简单和快速的。
Abstract Iron oxide nanoparticles have been used for many years as clinical applications. We have developed a rapid immunoaffinity isolation method of CD4+T cells from a mixed cell population of human blood using iron oxide nanoparticles. Anti CD4-antibody has been attached to iron oxide nanoparticles after its surface modification. The antibody tagged iron oxide nanoparticle beads are simply incubated with the mixed cell population of human blood and CD4+T cells are purified using an external magnetic field. The purification level was checked by fluorescence microscopy and flow cytometry. The purified CD4+T cells were digested with trypsin with different time periods and the products were analyzed by MALDI-TOF mass spectrometry, without further fractionation or purification, to obtain its proteome pattern. A database search showed a number of peptide masses matched specific to T-cell peptide masses. These results indicate that iron oxide nanoparticles are useful for CD4+T cell purification, and mass spectrometry based proteolytic fingerprint is simple and swift for identifying putative surface biomarkers from the whole cell surfaces.