Document Detail

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.
MedLine Citation:
PMID:  17570377     Owner:  NLM     Status:  MEDLINE    
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 2.5x10(3) cells for one injection and would be applicable to the determination of NeuGc in cell therapy products.
Noritaka Hashii; Nana Kawasaki; Yukari Nakajima; Masashi Toyoda; Yoko Katagiri; Satsuki Itoh; Akira Harazono; Akihiro Umezawa; Teruhide Yamaguchi
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2007-05-25
Journal Detail:
Title:  Journal of chromatography. A     Volume:  1160     ISSN:  0021-9673     ISO Abbreviation:  J Chromatogr A     Publication Date:  2007 Aug 
Date Detail:
Created Date:  2007-07-30     Completed Date:  2007-09-27     Revised Date:  2009-01-15    
Medline Journal Info:
Nlm Unique ID:  9318488     Medline TA:  J Chromatogr A     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  263-9     Citation Subset:  IM    
Division of Biological Chemistry and Biologicals, National Institute of Health Sciences, 1-18-1 Kamiyouga, Setagaya-ku, Tokyo 158-8501, Japan.
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MeSH Terms
Cell Membrane / chemistry
Chromatography, Liquid / methods*
Fourier Analysis*
HL-60 Cells
Mass Spectrometry / methods*
N-Acetylneuraminic Acid / analysis,  chemistry
Nanotechnology / methods*
Neuraminic Acids / analysis*,  chemistry
Quality Control
Subcellular Fractions / chemistry
Tissue Therapy / standards*
Reg. No./Substance:
0/Neuraminic Acids; 1113-83-3/N-glycolylneuraminic acid; 131-48-6/N-Acetylneuraminic Acid

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