使用带有共价连接免疫球蛋白G的蛋白A柱，通过高效免疫亲和色谱法检测真菌碳水化合物抗原。

Detection of fungal carbohydrate antigens by high-performance immunoaffinity chromatography using a protein A column with covalently linked immunoglobulin G.

作者信息

De Ruiter G A, Smid P, Schols H A, Van Boom J H, Rombouts F M

机构信息

Department of Food Science, Wageningen Agricultural University, Netherlands.

出版信息

J Chromatogr. 1992 Dec 11;584(1):69-75. doi: 10.1016/0378-4347(92)80010-n.

DOI:10.1016/0378-4347(92)80010-n

PMID:1487517

Abstract

Fungal carbohydrate antigens were analysed by high-performance immunoaffinity chromatography (HPIAC) with immunoglobulin G (IgG) antibodies raised against extracellular polysaccharides of Mucor racemosus. The protein A-IgG complex was covalently bound with dimethyl pimelimidate, which enabled the use of strong acidic buffers to release the tightly bound antigens from the column. Prior to pulsed-amperometric detection, an anion-micromembrane suppressor was used to raise the pH of the effluent to above 12 without dilution. The HPIAC system provides a sophisticated method for the rapid and sensitive detection of antigenic oligomeric carbohydrates in biological samples and is proposed as an alternative to quantitative enzyme-linked immunosorbent assay techniques.

摘要

采用针对总状毛霉细胞外多糖产生的免疫球蛋白G（IgG）抗体，通过高效免疫亲和色谱法（HPIAC）分析真菌碳水化合物抗原。蛋白A-IgG复合物用庚二酸二甲酯共价结合，这使得能够使用强酸性缓冲液从柱上释放紧密结合的抗原。在脉冲安培检测之前，使用阴离子微膜抑制器将流出物的pH值提高到12以上而不进行稀释。HPIAC系统为生物样品中抗原性低聚糖的快速灵敏检测提供了一种精密方法，并被提议作为定量酶联免疫吸附测定技术的替代方法。