Guzman Norberto A, Blanc Timothy, Phillips Terry M
Biomarker Laboratory, Princeton Biochemicals, Inc., Princeton, NJ 08543, USA.
Electrophoresis. 2008 Aug;29(16):3259-78. doi: 10.1002/elps.200800058.
In the last few years, there has been a greater appreciation by the scientific community of how separation science has contributed to the advancement of biomedical research. Despite past contributions in facilitating several biomedical breakthroughs, separation sciences still urgently need the development of improved methods for the separation and detection of biological and chemical substances. In particular, the challenging task of quantifying small molecules and biomolecules, found in low abundance in complex matrices (e.g., serum), is a particular area in need of new high-efficiency techniques. The tandem or on-line coupling of highly selective antibody capture agents with the high-resolving power of CE is being recognized as a powerful analytical tool for the enrichment and quantification of ultra-low abundance analytes in complex matrices. This development will have a significant impact on the identification and characterization of many putative biomarkers and on biomedical research in general. Immunoaffinity CE (IACE) technology is rapidly emerging as the most promising method for the analysis of low-abundance biomarkers; its power comes from a three-step procedure: (i) bioselective adsorption and (ii) subsequent recovery of compounds from an immobilized affinity ligand followed by (iii) separation of the enriched compounds. This technology is highly suited to automation and can be engineered to as a multiplex instrument capable of routinely performing hundreds of assays per day. Furthermore, a significant enhancement in sensitivity can be achieved for the purified and enriched affinity targeted analytes. Thus, a compound that exists in a complex biological matrix at a concentration far below its LOD is easily brought to well within its range of quantification. The present review summarizes several applications of IACE, as well as a chronological description of the improvements made in the fabrication of the analyte concentrator-microreactor device leading to the development of a multidimensional biomarker analyzer.
在过去几年中，科学界对分离科学如何推动生物医学研究的进展有了更深刻的认识。尽管分离科学在促成多项生物医学突破方面过去做出了贡献，但仍迫切需要开发改进的生物和化学物质分离与检测方法。特别是，在复杂基质（如血清）中低丰度存在的小分子和生物分子的定量这一具有挑战性的任务，是一个尤其需要新型高效技术的领域。高选择性抗体捕获剂与毛细管电泳的高分辨率进行串联或在线联用，正被公认为是用于富集和定量复杂基质中超低丰度分析物的强大分析工具。这一进展将对许多假定生物标志物的鉴定和表征以及总体生物医学研究产生重大影响。免疫亲和毛细管电泳（IACE）技术正迅速成为分析低丰度生物标志物最有前景的方法；其优势源于一个三步程序：（i）生物选择性吸附，（ii）随后从固定化亲和配体中回收化合物，接着（iii）分离富集的化合物。该技术非常适合自动化，并且可以设计成能够每天常规进行数百次检测的多重仪器。此外，对于纯化和富集的亲和靶向分析物，可以实现灵敏度的显著提高。因此，一种存在于复杂生物基质中、浓度远低于其检测限的化合物能够很容易地被带到其定量范围内。本综述总结了IACE的几种应用，以及对分析物浓缩器 - 微反应器装置制造过程中所做改进的按时间顺序的描述，这些改进促成了多维生物标志物分析仪的开发。
