Accurate quantification of beta-hexosaminidase released from laboratory of allergic diseases 2 cells via liquid chromatography tandem mass spectrometry method

作者：Lv, Yanni; Fu, Jia; Jia, Qianqian; Che, Delu; Lin, Yuanyuan; Han, Shengli^*; He, Langchong

来源：Journal of Chromatography A, 2018, 1578: 106-111.

DOI：10.1016/j.chroma.2018.09.059

• LC-MS/MS
• Anaphylactoid reaction
• LAD2 cell
• beta-Hexosaminidase
• Mass spectrometry

beta-Hexosaminidase is one of the enzymes that is secreted from mast cells via antigen-induced degranulation and has frequently been used as an indicator of anaphylactic reactions. The main method for determining beta-hexosaminidase is indirect "substrate-based" and shows limitations. Hence development of an accurate detecting method is particularly important and urgently needed. In this study we developed a liquid chromatography tandem mass spectrometry (LC-MS/MS) method for measuring beta-hexosaminidase. Laboratory of allergic diseases 2 (LAD2) cells were stimulated with compound 48/80 (C48/80), the supernatant was collected and subjected to in-solution protein digestion; the obtained peptides were desalted, concentrated, separated and analyzed with an LC-tandem MS instrument. A peptide with the sequence "LAPGTIVEVWK" was selected for quantitative analysis, and four other peptides for qualitative research. The time-effect relationship curve was studied, and the results of the LC-MS/MS method were found to be almost consistent with those obtained via the conventional method. The method was then employed to measure beta-hexosaminidase released from LAD2 cells stimulated with potential allergens, and the results showed that it can be applied to determine the potential allergenicity of drugs. This new method showed good specificity, high sensitivity and a wide application range. It could be used to evaluate allergic reactions, providing a guide for medication safety during clinical testing.

• 出版日期2018-11-30
• 单位西安交通大学