新型血清变异转甲状腺素蛋白的简便快速分析方法：遗传性转甲状腺素蛋白淀粉样变性的直接 MALDI 筛查方法。
New simple and quick method to analyze serum variant transthyretins: direct MALDI method for the screening of hereditary transthyretin amyloidosis.
机构信息
Department of Neurology, Graduate School of Medical Sciences, Kumamoto University, 1-1-1 Honjo, Kumamoto, 860-0811, Japan.
Department of Morphological and Physiological Sciences, Graduate School of Health Sciences, Kumamoto University, 4-24-1 Kuhonji, Kumamoto, 862-0976, Japan.
出版信息
Orphanet J Rare Dis. 2019 May 27;14(1):116. doi: 10.1186/s13023-019-1100-y.
BACKGROUND
Hereditary transthyretin amyloidosis (ATTRv amyloidosis) is caused by a variant transthyretin (TTR), which is a serum protein secreted by the liver. Mass spectrometry (MS) is a useful tool that can detect variant TTRs in serum samples from patients with ATTRv amyloidosis. We previously reported several mass spectrometric methods to detect variant TTRs in serum samples. Those methods require cumbersome immunoprecipitation with anti-TTR antibodies and significant time to analyze the variant TTRs. In our study here, we developed a new simple and quick method to detect variant TTRs in serum samples by means of matrix-assisted laser desorption-ionization time-of-flight (MALDI-TOF) MS without immunoprecipitation (direct MALDI).
METHODS
By using direct MALDI, we analyzed 288 serum samples obtained from patients who were clinically suspected having amyloidosis to investigate the usefulness of this direct MALDI method to detect variant TTRs in serum samples.
RESULTS
The method completed the process within 30 min. We successfully identified variant TTRs in serum samples from patients, except for a few patients with TTR Glu61Lys and Glu89Gln mutations because of the small mass shift of those variant TTRs from wild-type TTR. We also found that the mass shifts of variant TTRs measured by direct MALDI corresponded to theoretical mass changes.
CONCLUSION
Our results suggest that the direct MALDI method is useful for the screening of ATTRv amyloidosis.
背景
遗传性转甲状腺素蛋白淀粉样变性（ATTRv 淀粉样变性）是由变异转甲状腺素蛋白（TTR）引起的，TTR 是一种由肝脏分泌的血清蛋白。质谱（MS）是一种有用的工具，可以检测到 ATTRv 淀粉样变性患者血清样本中的变异 TTR。我们之前报道了几种用于检测血清样本中变异 TTR 的质谱方法。这些方法需要用抗 TTR 抗体进行繁琐的免疫沉淀，并且需要大量时间来分析变异 TTR。在我们的这项研究中，我们开发了一种新的简单快速的方法，通过基质辅助激光解吸电离飞行时间（MALDI-TOF）MS 而无需免疫沉淀（直接 MALDI）来检测血清样本中的变异 TTR。
方法
通过使用直接 MALDI，我们分析了 288 份来自临床疑似患有淀粉样变性的患者的血清样本，以研究该直接 MALDI 方法在检测血清样本中变异 TTR 的有用性。
结果
该方法在 30 分钟内完成了整个过程。我们成功地在患者的血清样本中识别出了变异 TTR，除了少数 TTR Glu61Lys 和 Glu89Gln 突变的患者，因为这些变异 TTR 与野生型 TTR 的质量偏移较小。我们还发现，直接 MALDI 测量的变异 TTR 的质量偏移与理论质量变化相对应。
结论
我们的结果表明，直接 MALDI 方法可用于 ATTRv 淀粉样变性的筛查。
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