TY - JOUR
T1 - Accurate quantification of β-hexosaminidase released from laboratory of allergic diseases 2 cells via liquid chromatography tandem mass spectrometry method
AU - Lv, Yanni
AU - Fu, Jia
AU - Jia, Qianqian
AU - Che, Delu
AU - Lin, Yuanyuan
AU - Han, Shengli
AU - He, Langchong
N1 - Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2018/11/30
Y1 - 2018/11/30
N2 - β-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 β-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 β-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 β-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.
AB - β-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 β-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 β-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 β-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.
KW - Anaphylactoid reaction
KW - LAD2 cell
KW - LC–MS/MS
KW - Mass spectrometry
KW - β-Hexosaminidase
UR - https://www.scopus.com/pages/publications/85054384601
U2 - 10.1016/j.chroma.2018.09.059
DO - 10.1016/j.chroma.2018.09.059
M3 - 文章
C2 - 30297235
AN - SCOPUS:85054384601
SN - 0021-9673
VL - 1578
SP - 106
EP - 111
JO - Journal of Chromatography A
JF - Journal of Chromatography A
ER -