在线免疫亲和固相萃取毛细管电泳-质谱法分析血清转甲状腺素蛋白
On-Line Immunoaffinity Solid-Phase Extraction Capillary Electrophoresis-Mass Spectrometry for the Analysis of Serum Transthyretin.
作者信息
Pero-Gascon Roger, Pont Laura, Sanz-Nebot Victoria, Benavente Fernando
机构信息
Department of Chemical Engineering and Analytical Chemistry, Institute for Research on Nutrition and Food Safety (INSA·UB), University of Barcelona, Barcelona, Spain.
出版信息
Methods Mol Biol. 2019;1972:57-76. doi: 10.1007/978-1-4939-9213-3_5.
The analysis of low abundant proteins in biological fluids by capillary electrophoresis (CE) is particularly problematic due to the typically poor concentration limits of detection of microscale separation techniques. Another important issue is sample matrix complexity that requires an appropriate cleanup. Here, we describe an on-line immunoaffinity solid-phase extraction capillary electrophoresis-mass spectrometry (IA-SPE-CE-MS) method for the immunoextraction, preconcentration, separation, detection, and characterization of serum transthyretin (TTR). TTR is a protein biomarker related to diverse types of amyloidosis, such as familial amyloidotic polyneuropathy type I (FAP-I), which is the most common hereditary systemic amyloidosis.
由于微量分离技术通常较差的检测浓度限，通过毛细管电泳（CE）分析生物流体中的低丰度蛋白质特别具有挑战性。另一个重要问题是样品基质的复杂性，这需要进行适当的净化处理。在此，我们描述了一种在线免疫亲和固相萃取毛细管电泳-质谱联用（IA-SPE-CE-MS）方法，用于血清转甲状腺素蛋白（TTR）的免疫提取、预浓缩、分离、检测和表征。TTR是一种与多种类型淀粉样变性相关的蛋白质生物标志物，如I型家族性淀粉样多神经病（FAP-I），它是最常见的遗传性全身性淀粉样变性。
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