Smarter staining: New biocatalytic systems for multiplexing histological staining

背景：组织病理学是许多医学实验室的主力，可用于疾病诊断、药物开发和生物学研究。组织学染色是这一过程的重要工具，它提供了有价值的对比度，并直观地识别了样品中的微观特征。由于这一过程既要稳健，又要可重复，这一点至关重要，创新受到了限制，只采用了极少数的技术。这些技术中最主要的是染色过程，它使用辣根过氧化物酶(HRP)催化二氨基联苯胺(DAB)的氧化，产生不溶于水的棕色染色。当与抗体识别系统相结合时，这可以提供生物组织样本上任何单个目标的地图。随着“个人医学”的出现，多重染色将在组织病理学方面提供重要的进步，因为可以从许多患者样本中产生信息量，这是组织受限的一个特别重要的特征。目前，多重作用仅限于使用连接到两种不同酶/色原系统的小鼠和兔抗体：HRP/DAB组合和正交碱性磷酸酶(AP)/坚固红组合。在本项目中：将探索一种创新的多重作用方法，该方法依赖于新的聚合物结合的生物催化剂/前着色原底物组合的开发，其中前着色原底物的生物催化周转可用于多重染色。

Background: Histopathology is the workhorse of many medical laboratories, and can be used for diseasediagnosis, drug development and biological research. Histological staining is an essential tool for this process,providing valuable contrast and visually identifying microscopic features in samples. Since it is essential thatthis process is both robust and repeatable, innovation has been limited and only a very few technologies havebeen employed. Chief amongst these technologies are staining process which use horse radish peroxidase(HRP) to catalyze the oxidation of the chromogen diaminobenzidine (DAB) to give an insoluble brown stain.When coupled to an antibody recognition system this can provide a map of any individual target on thebiological tissue sample. With the advent of "personal medicine", multiplexing staining would provide animportant advance in histopathology in terms of the amount of information which could be generated fromany one patient sample, a particularly important feature where tissue is limited. At the moment multiplexingis limited to the use of mouse and rabbit antibodies linked to two different enzyme/chromogen systems: theHRP/DAB combination; and an orthogonal alkaline phosphatase (AP)/Fast Red combination.In this project: an innovative approach to multiplexing will be explored which relies upon the development ofnew polymer-bound biocatalyst /pro-chromogen substrate combinations, in which biocatalytic turnover ofthe pro-chromogen substrate can be used to multiplex staining.