Detection of transthyretin variants using immunoprecipitation and matrix-assisted laser desorption/ionization bioreactive probes: a clinical application of mass spectrometry.
Detection of transthyretin variants using immunoprecipitation and matrix-assisted laser desorption/ionization bioreactive probes: a clinical application of mass spectrometry.
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使用免疫沉淀和基质辅助激光解吸/电离生物反应探针检测运甲状腺素蛋白变体：质谱的临床应用。
DOI：
10.1016/s1044-0305(99)00136-1
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发表时间：
2000
影响因子：
3.2
通讯作者：
Costello,CE
中科院分区：
文献类型：
--
作者：
Théberge,R;Connors,LH;Skinner,M;Costello,CE
在我们不断努力开发基于质谱学的方法来检测和表征转甲状腺素变异体的过程中，我们试图使用含有固定化胰酶的基质辅助激光解吸/电离(MALDI)生物反应探针进行筛选。这些设备显示出很好的诊断潜力，可以作为临床筛查工具来检测TTR中的氨基酸替换。MALDI探针允许在探针上生成胰酶消化。随后对探针上的消化进行质量分析，得到了肽图。这种方法的固有优势包括大大缩短了消化时间(几分钟比几小时)，不需要自溶产品，最大限度地减少了样品处理，因此样品损失最小。另一个优点是，由于不会发生样品转移，因此减少了疏水性多肽丢失的机会。该方法可用于通过免疫沉淀法从患者血清中分离出TTR变异体的初步筛选。该方法也应适用于其他蛋白质，并适合于自动化。
In our continuing efforts to develop mass spectrometry-based methods for transthyretin (TTR) variant detection and characterization, we have sought to use matrix-assisted laser desorption/ionization (MALDI) bioreactive probes incorporating immobilized trypsin for screening purposes. These devices show good diagnostic potential as a clinical screening tool to detect amino acid substitutions in TTR. MALDI probes allow the on-probe generation of tryptic digests. The subsequent mass analysis of the on-probe digest yields the peptide map. The inherent advantages of this method include considerably reduced digestion times (minutes vs. hours), absence of autolysis products, minimized sample handling, and hence minimal sample loss. A further advantage is that the opportunity for loss of hydrophobic peptides is reduced because no sample transfer occurs. The method can be applied as a preliminary screen for TTR variants where TTR is isolated from patient serum through immunoprecipitation. This method should also be applicable to other proteins and suitable for automation.