细胞间黏附分子1的MRM-MS测定方法建立与优化

黄东萍; 李洪涛; 曾小云; 龙静铧; 臧宁; 周怡; 仇小强

doi:10.11847/zgggws1118782

细胞间黏附分子1的MRM-MS测定方法建立与优化

Establishment and optimization of a MRM-MS method for detection of intercellular adhesion molecule 1

摘要

摘要:
目的建立并优化室间隔缺损潜在生物标记物细胞间黏附分子1（ICAM-1）MRM-MS测定方法。
方法借助UniProt数据库和辅助软件Skyline建立ICAM-1多重反应监测质谱定量（MRM-MS）方法，进一步筛选其各肽段及母子离子对，并通过改变MS的碰撞能量CE值和去簇电压DP值进一步优化其MRM质谱扫描方法。
结果从ICAM-1的11个肽段88对离子对中优选出3个肽段9对离子对作为ICAM-1的定量依据。采用优化后的MRM-MS进行检测，10个血清样本中ICAM-1蛋白含量为（318.40 ± 97.20）ng/mL，与经酶联免疫法测定的结果（330.30 ± 130.24）ng/mL相比差异无统计学意义（P > 0.05）。
结论借助UniProt数据库和辅助软件Skyline，经过多个循环可以建立并优化ICAM-1蛋白的MRM-MS测定方法。

Abstract:
Objective To establish and optimize a multiple reaction monitoring-mass spectrometry (MRM-MS) method for detection of intercellular adhesion molecule 1 (ICAM-1), which is a potential biomarker for ventricular septal defect.
Methods By means of UniProt database and the Skyline software we established a MRM-MS method to detect ICAM-1; then we screened the peptides and the ion pair of ICAM-1 and changed the collision energy value and declustering potential voltage to optimize the MRM-MS method.
Results Three peptides and 9 ion pairs were selected from 11 peptides and 88 ion pairs of ICAM-1 as the basis of quantitative determination. Detected with the optimized MRM-MS, the content of ICAM-1 protein in 10 serum samples was 318.40 ± 97.20 ng/ml, which was not significantly different from the result (330.30 ± 130.24 ng/ml) measured with enzyme-linked immunoassay (P > 0.05).
Conclusion By means of UniProt database and the Skyline software and several procedures, we established and optimized a MRM-MS method for the detection of ICAM-1.

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