Study on the quality control of cell therapy products Determination of N-glycolylneuraminic acid incorporated into human cells by nano-flow liquid chromatography Fourier transformation ion cyclotron mass spectrometry
Study on the quality control of cell therapy products Determination of N-glycolylneuraminic acid incorporated into human cells by nano-flow liquid chromatography Fourier transformation ion cyclotron mass spectrometry
复制标题
DOI：
10.1016/j.chroma.2007.05.062
复制
发表时间：
2007-08-10
影响因子：
4.1
通讯作者：
Yamaguchi, Teruhide
中科院分区：
文献类型：
--
作者：
Hashii, Noritaka;Kawasaki, Nana;Yamaguchi, Teruhide
n -糖基神经氨酸（NeuGc）是一种酸性九碳糖，由牛和老鼠等几种动物产生。由于人类细胞不能合成NeuGc，因此被认为在人类中具有免疫原性。最近，在异种血清和细胞培养的人胚胎干细胞中有NeuGc污染的报道，这表明NeuGc可能会损害细胞治疗产品的有效性和安全性。唾液酸用1,2-二氨基-4,5-亚甲基二氧苯（DMB）衍生化，然后用液相色谱/质谱（LC/MS）和液相色谱/串联质谱（LC/MS/MS）测定；然而，有限的细胞治疗产品的可用性需要更敏感和特定的方法进行质量检测。本文研究了纳米流液相色谱/傅里叶变换离子回旋共振质谱（nanoLC/FTMS）和纳米olc /MS/MS在低飞摩尔水平下对neugc的特异性测定。用我们的方法，我们发现用人血清培养的人细胞系（HL-60RG细胞）被NeuGc污染。我们的方法一次注射仅需2.5 × 10(3)个细胞，适用于细胞治疗产品中NeuGc的测定。(c) 2007 Elsevier B.V.版权所有
N-Glycolylneuraminic acid (NeuGc), an acidic nine-carbon sugar, is produced in several animals, such as cattle and mice. Since human cells cannot synthesize NeuGc, it is considered to be immunogenic in humans. Recently, NeuGc contamination was reported in human embryonic stem cells cultured with xenogeneic serum and cells, suggesting that possibly NeuGc may harm the efficacy and safety of cell therapy products. Sialic acids have been determined by derivatization with 1,2-diamino-4,5-methylenedioxybenzene (DMB) followed by liquid chromatography/mass spectrometry (LC/MS) and liquid chromatography/tandem mass spectrometry (LC/MS/MS); however, the limited availability of cell therapy products requires more sensitive and specific methods for the quality test. Here we studied the use of nano-flow liquid chromatography/Fourier transformation ion cyclotron resonance mass spectrometry (nanoLC/FTMS) and nanoLC/MS/MS for NeuGc-specific determination at a low femtomole level. Using our method, we found NeuGc contamination of the human cell line (HL-60RG cells) cultured with human serum. Our method needs only 2.5 x 10(3) cells for one injection and would be applicable to the determination of NeuGc in cell therapy products. (c) 2007 Elsevier B.V. All rights reserved.