Liquid Chromatography-High Resolution Mass Spectrometry Analysis of Platelet Frataxin as a Protein Biomarker for the Rare Disease Friedreich’s Ataxia,Analytical Chemistry

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Liquid Chromatography-High Resolution Mass Spectrometry Analysis of Platelet Frataxin as a Protein Biomarker for the Rare Disease Friedreich’s Ataxia
Analytical Chemistry ( IF 6.7 ) Pub Date : 2018-01-11 00:00:00 , DOI: 10.1021/acs.analchem.7b04590
Lili Guo _{1,

2} , Qingqing Wang _{1,

2} , Liwei Weng ₁ , Lauren A Hauser _{2,

3,

4} , Cassandra J Strawser _{2,

3,

4} , Agostinho G Rocha ₅ , Andrew Dancis ₅ , Clementina Mesaros _{1,

2} , David R Lynch _{2,

3,

4} , Ian A Blair _{1,

2}

Affiliation

Friedreich’s ataxia (FA) is an autosomal recessive disease caused by an intronic GAA triplet expansion in the FXN gene, leading to reduced expression of the mitochondrial protein frataxin. FA is estimated to affect 1 in 50 000 with a mean age of death in the fourth decade of life. There are no approved treatments for FA, although experimental approaches, which involve up-regulation or replacement of frataxin protein, are being tested. Frataxin is undetectable in serum or plasma, and whole blood cannot be used because it is present in long-lived erythrocytes. Therefore, an assay was developed for analyzing frataxin in platelets, which have a half-life of 10 days. The assay is based on stable isotope dilution immunopurification two-dimensional nano-ultra high performance liquid chromatography/parallel reaction monitoring/mass spectrometry. The lower limit of quantification was 0.078 pg frataxin/μg protein, and the assay had 100% sensitivity and specificity for discriminating between controls and FA cases. The mean levels of control and FA platelet frataxin were 9.4 ± 2.6 and 2.4 ± 0.6 pg/μg protein, respectively. The assay should make it possible to rigorously monitor the effects of therapeutic interventions on frataxin expression in this devastating disease.

中文翻译：

液相色谱-高分辨率质谱分析血小板 Frataxin 作为罕见疾病弗里德赖希共济失调的蛋白质生物标志物

弗里德赖希共济失调 (FA) 是一种常染色体隐性遗传疾病，由FXN基因中的内含子 GAA 三联体扩增引起，导致线粒体蛋白 frataxin 表达减少。据估计，FA 会影响五万分之一的人，平均死亡年龄在四十岁左右。尽管正在测试涉及上调或替换 frataxin 蛋白的实验方法，但尚无批准的 FA 治疗方法。 Frataxin 在血清或血浆中检测不到，并且不能使用全血，因为它存在于长寿命红细胞中。因此，开发了一种分析血小板中 frataxin 的方法，其半衰期为 10 天。该测定基于稳定同位素稀释免疫纯化二维纳米超高效液相色谱/平行反应监测/质谱。定量下限为 0.078 pg frataxin/μg 蛋白，该测定对于区分对照和 FA 病例具有 100% 的敏感性和特异性。对照和 FA 血小板 frataxin 的平均水平分别为 9.4 ± 2.6 和 2.4 ± 0.6 pg/μg 蛋白质。该测定应该能够严格监测治疗干预措施对这种毁灭性疾病中 frataxin 表达的影响。

更新日期：2018-01-11

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