凝集素的纯化、定量和功能分析。

Purification, Quantification, and Functional Analysis of Collectins.

机构信息

Department of Clinical Nutrition, Rakuno Gakuen University, Hokkaido, Japan.

Department of Medicine and Physiology, Rakuno Gakuen University, Hokkaido, Japan.

出版信息

Methods Mol Biol. 2020;2132:99-109. doi: 10.1007/978-1-0716-0430-4_10.

DOI:10.1007/978-1-0716-0430-4_10

PMID:32306318

Abstract

Native and recombinant collectins are purified by using mannan-agarose and an anti-collectin antibody column. The use of sandwich enzyme-linked immunosorbent assay (ELISA) with two antibodies against human mannan-binding lectin (MBL) enables elucidation of the collectin concentration in the blood, serum, and plasma. The collectin sugar specificity is demonstrated by determining the concentration of saccharide required to inhibit sugar binding by 50% in a saccharide-binding assay. Biological analyses including the complement-dependent hemolysis test and several other methods are used to evaluate collectin.

摘要

天然和重组凝集素可通过甘露聚糖琼脂和抗凝集素抗体柱进行纯化。夹心酶联免疫吸附试验（ELISA）使用两种针对人甘露聚糖结合凝集素（MBL）的抗体，可阐明血液、血清和血浆中的凝集素浓度。通过测定糖结合抑制试验中抑制糖结合 50%所需的糖浓度来确定凝集素的糖特异性。生物学分析包括补体依赖性溶血试验和其他几种方法用于评估凝集素。