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在非均相介质中使用带有扩展微床吸附柱的流动注射系统进行结合测定。

Binding assays in heterogeneous media using a flow injection system with an expanded micro-bed adsorption column.

作者信息

Mattiasson B, Nandakumar M P

机构信息

Department of Biotechnology, Lund University, Sweden.

出版信息

Bioseparation. 1999;8(1-5):237-45.

Abstract

Competitive binding assays have been performed in flow injection systems. To further increase the versatility of the system, and to enable it to deal with samples containing particulate matter, the adsorption step was designed as an expanded bed column. Immunochemical quantification of human serum albumin was chosen as a model system to use for the development of the technology. A competitive ELISA was set up using peroxidase labelled HSA as competing ligand. The introduction of the expanded bed immunosorption column made the system tolerant to samples containing suspended particulate matter. The analytical outcome is very similar to that from the packed bed system even though more time is required for each assay cycle. The capability of the system was tested by addition of increasing amounts of yeast cells. The results clearly indicate that the system is suitable e.g. for process monitoring of fermentations.

摘要

竞争结合测定已在流动注射系统中进行。为了进一步提高该系统的通用性,并使其能够处理含有颗粒物的样品,吸附步骤设计为膨胀床柱。选择人血清白蛋白的免疫化学定量作为用于该技术开发的模型系统。使用过氧化物酶标记的人血清白蛋白作为竞争配体建立了竞争性酶联免疫吸附测定。膨胀床免疫吸附柱的引入使该系统能够耐受含有悬浮颗粒物的样品。即使每个测定周期需要更多时间,分析结果与填充床系统的结果非常相似。通过添加越来越多的酵母细胞来测试该系统的性能。结果清楚地表明该系统适用于例如发酵过程监测。

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