Department of Genetics and Pathology, Uppsala University, Uppsala, Sweden.
J Intern Med. 2010 Sep;268(3):232-45. doi: 10.1111/j.1365-2796.2010.02256.x.
The ability to detect very low levels of expressed proteins has enormous potential for early diagnostics and intervention at curable stages of disease. An extended range of targets such as interacting or post-translationally modified proteins can further improve the potential for diagnostics and patient stratification, and for monitoring response to treatment. These are critical building blocks for personalized treatment strategies to manage disease. The past few decades have seen a remarkably improved understanding of the molecular basis of disease in general, and of tumour formation and progression in particular. This accumulated knowledge creates opportunities to develop drugs that specifically target molecules or molecular complexes critical for survival and expansion of tumour cells. However, tumours are highly variable between patients, necessitating the development of diagnostic tools to individualize treatment through parallel analysis of sets of biomarkers. The proximity ligation assay (PLA) can address many of the requirements for advanced molecular analysis. The method builds on the principle that recognition of target proteins by two, three or more antibodies can bring in proximity DNA strands attached to the antibodies. The DNA strands can then participate in ligation reactions, giving rise to molecules that are amplified for highly sensitive detection. PLA is particularly well suited for sensitive, specific and multiplexed analysis of protein expression, post-translational modifications and protein-protein interactions. The analysis of this extended range of biomarkers will prove critical for the development and implementation of personalized medicine.
检测表达蛋白的极低水平的能力具有在疾病可治愈阶段进行早期诊断和干预的巨大潜力。更广泛的目标，如相互作用或翻译后修饰的蛋白质，可以进一步提高诊断和患者分层的潜力，并监测对治疗的反应。这些是管理疾病的个性化治疗策略的关键组成部分。在过去的几十年中，人们对疾病的分子基础有了显著的深入了解，特别是对肿瘤的形成和发展有了更深入的了解。这些积累的知识为开发专门针对肿瘤细胞存活和扩张至关重要的分子或分子复合物的药物创造了机会。然而，肿瘤在患者之间具有高度的可变性，因此需要开发诊断工具，通过平行分析一组生物标志物来实现个体化治疗。邻近连接检测（PLA）可以满足高级分子分析的许多要求。该方法基于以下原理：通过两个、三个或更多抗体识别靶蛋白，可以使附着在抗体上的 DNA 链接近。然后，这些 DNA 链可以参与连接反应，产生可用于高度敏感检测的扩增分子。PLA 特别适合用于检测蛋白质表达、翻译后修饰和蛋白质-蛋白质相互作用的灵敏、特异和多重分析。对这种扩展范围的生物标志物的分析将证明对个性化药物的开发和实施至关重要。
