摘要
目的建立一种毛细管电泳(CE)与基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/MS)联用测定血清转甲状腺素蛋白(TTR)化学修饰的方法。方法用酶联免疫吸附法(ELISA)测定20例老年性淀粉样变(SSA)患者及20名健康体检者血清TTR浓度;CE法分离血清TTR并利用MALDI-TOF/MS测定TTR的化学修饰。结果健康对照组及SSA组血清TTR浓度分别为0.25±0.04(、0.26±0.03)g/L,2组间差异无统计学意义(P>0.05);MALDI-TOF/MS分析共发现4个TTR化学修饰峰,质/荷比(m/z)分别为:13 750.86±1.48、13 830.63±2.761、3 871.70±2.70、13 919.00±14.77;分别对应未修饰-TTR、磺化-TTR、半胱氨酸-TTR和半胱氨酸甘氨酸-TTR。结论 CE法分离血清TTR能够直接利用MALDI-TOF-MS测定其化学修饰,可用于临床检测血清TTR的化学修饰。
Objective To establish a combined method for analysis of serum transthyretin(TTR) chemical modification using capillary electrophoresis(CE) with matrix-assisted laser desorption ionization-time-of-flight mass spectrometry(MALDI-TOF/MS) analysis.Methods Enzyme-linked immunosorbent assay(ELISA) was used to determine serum TTR in 20 senile systemic amyloidosis(SSA) patients and 20 healthy subjects.Serum TTR was separated by CE.Determination of TTR chemical modification was analyzed by MALDI-TOF/MS.Results The levels of serum TTR was(0.25±0.04)g/L in control group and(0.26±0.03)g/L in SSA patients,and there was no significant difference between control group and SSA patients(P0.05).The chemical modification of TTR found 4 TTR chemical modification peaks,as native TTR [mass-to-charge ratio(m/z): 13 750.86±1.48],sul-TTR(m/z ratio: 13 830.63±2.76),cys-TTR(m/z ratio: 13 871.70±2.70) and cysgly-TTR(m/z ratio: 13 919.00±14.77) by MALDI-TOF/MS analysis.Conclusions The combined method of CE with MALDI-TOF/MS analysis can be used directly to determine the chemical modification of TTR.
出处
《检验医学》
CAS
2012年第3期185-188,共4页
Laboratory Medicine
关键词
转甲状腺素蛋白
毛细管电泳
质谱
化学修饰
Transthyretin
Capillary electrophoresis
Mass spectrometry
Chemical modification