Borén Mats
Denator AB, Dag Hammarskjöldsv. 10A, 753 81, Uppsala, Sweden,
Methods Mol Biol. 2015;1295:21-32. doi: 10.1007/978-1-4939-2550-6_2.
Due to post-sampling changes, caused by residual enzyme activity in the sample, levels of analytes can change from their in vivo levels so that they no longer accurately reflect conditions in the living system. The Stabilizor(™) system accomplishes elimination of enzyme activity through heat-induced denaturation of enzymes by permanently altering the 3D protein structure of the enzymes. Heat stabilization can be introduced in the workflow either directly after sampling, with the instrument just next to where the sample is taken, or prior to sample homogenization and extraction, when samples are heat denatured directly from a frozen state. Initially, heat stabilization was developed to enable mass spectrometric analysis of neuropeptides. Heat stabilization has since been further developed and applied to a range of samples and downstream protein analysis techniques such as western blot, 2D gels and phosphorylation analysis with LC-MS.
由于样本中残留的酶活性导致采样后发生变化，分析物的水平可能会与其体内水平不同，从而无法再准确反映生物系统中的状况。Stabilizor(™)系统通过热诱导酶变性来消除酶活性，即永久性改变酶的三维蛋白质结构。热稳定化可以在采样后直接在仪器旁紧邻采样点的位置引入工作流程，也可以在样品匀浆和提取之前，将样品直接从冷冻状态进行热变性时引入。最初，热稳定化技术是为了实现神经肽的质谱分析而开发的。此后，热稳定化技术得到了进一步发展，并应用于一系列样品和下游蛋白质分析技术，如蛋白质印迹、二维凝胶电泳以及液相色谱-质谱联用的磷酸化分析。
