Guzman N A
Bioanalytical Drug Metabolism, The R.W. Johnson Pharmaceutical Research Institute, Raritan, NJ 08896, USA.
J Chromatogr B Biomed Sci Appl. 2000 Dec 1;749(2):197-213. doi: 10.1016/s0378-4347(00)00410-2.
The need for urgent diagnoses has propelled the development of automated analyses that can be performed in a short time at reasonable cost. One such method is immunoaffinity capillary electrophoresis. This emerging hybrid technology employs two powerful techniques coupled on-line for the direct and rapid determination of analytes present in biological fluids. The first technique, immunoaffinity, is used for the selective extraction of a molecule present in a complex matrix, utilizing a microscale-format chamber affinity device. An analyte (affinity target) present in serum or urine is captured by an immobilized molecular recognition antibody molecule (affinity ligand) bound to a solid support constituent (glass beads or an appropriate porous structure) of a microchamber affinity device. The second technique, capillary electrophoresis, is used for the high-resolution analytical separation of the purified and concentrated affinity target material after elution from the microchamber affinity device. In this work, immunoaffinity capillary electrophoresis was developed for the identification and characterization of a single constituent of a complex matrix. Immunoreactive gonadotropin-releasing hormone was determined in serum and urine specimens derived from a normal individual and from a patient suffering from benign prostatic hyperplasia. Furthermore, the on-line immuno-separation system was coupled in tandem to mass spectrometry to obtain molecular mass information of the affinity isolated and CE separated neuropeptide. This hybrid immuno-analytical technology is simple, rapid, selective and sensitive. In addition, an attempt was also made to characterize other urinary constituents by CE-MS that may lead to marker activity in the urine of the diseased subject. The hyphenation of analytical techniques has proved valuable in enhancing their individual features. The future of bioanalysis using miniaturized affinity systems is discussed in this paper.
对紧急诊断的需求推动了自动化分析技术的发展，这类技术能够在短时间内以合理成本进行检测。免疫亲和毛细管电泳就是其中一种方法。这种新兴的混合技术在线联用了两种强大的技术，用于直接快速测定生物体液中的分析物。第一种技术是免疫亲和，它利用微尺度腔室亲和装置，对复杂基质中存在的分子进行选择性提取。血清或尿液中的分析物（亲和靶标）被固定在微腔室亲和装置固体支持成分（玻璃珠或合适的多孔结构）上的分子识别抗体分子（亲和配体）捕获。第二种技术是毛细管电泳，用于对从微腔室亲和装置洗脱后的纯化浓缩亲和靶标物质进行高分辨率分析分离。在这项工作中，开发了免疫亲和毛细管电泳用于鉴定和表征复杂基质中的单一成分。测定了来自正常个体和良性前列腺增生患者的血清和尿液样本中的免疫反应性促性腺激素释放激素。此外，在线免疫分离系统与质谱串联联用，以获取亲和分离和毛细管电泳分离后的神经肽的分子量信息。这种混合免疫分析技术简单、快速、具有选择性且灵敏。此外，还尝试通过毛细管电泳 - 质谱联用对其他尿液成分进行表征，这些成分可能在患病个体尿液中具有标志物活性。分析技术的联用已证明在增强其各自特性方面具有价值。本文讨论了使用小型化亲和系统进行生物分析的未来发展。
