Traditional immunoprobing techniques like Western blot continue to play a crucial role in the discovery and validation of biomarkers. This technique suffers from several limitations that affect reproducibility and feasibility for large-scale studies. Modern immunoprobing techniques have addressed several of these limitations. Here we contrast the use of Western blot and an automated capillary nano-immunoassay (CNIA), Wes™. We provide evidence highlighting the methodological advantages of Wes™ over Western blot in the validation of a novel biomarker, Calcium-binding protein and spermatid-associated 1 (hCABS1). While Wes™ offers a faster, more consistent approach with lower requirements for sample and antibody volumes, variations in expected molecular weights and computational algorithms used to analyze the data must receive careful consideration and assessment. Our data suggests that CNIA approaches are likely to positively impact biomarker discovery and validation.

中文翻译：

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一种使用自动毛细管纳米免疫测定平台（Wes™）研究唾液中蛋白质生物标志物的方法。

传统的免疫探测技术（如蛋白质印迹）在生物标志物的发现和验证中继续发挥关键作用。这项技术有几个局限性，影响了大规模研究的可重复性和可行性。现代的免疫探测技术已经解决了其中一些局限性。在这里，我们对比了蛋白质印迹和自动毛细管纳米免疫测定（CNIA）Wes™的使用。我们提供了证据，证明了在验证新型生物标志物，钙结合蛋白和与精子相关的1（hCABS1）方面，Wes™优于蛋白质印迹的方法学优势。尽管Wes™提供了一种更快，更一致的方法，但对样品和抗体量的要求却较低，预期分子量的变化和用于分析数据的计算算法必须经过仔细考虑和评估。我们的数据表明，CNIA方法可能会对生物标志物的发现和验证产生积极影响。