通过单克隆抗体辅助免疫亲和色谱法纯化甲状腺过氧化物酶。

Purification of thyroid peroxidase by monoclonal antibody-assisted immunoaffinity chromatography.

作者信息

Nakagawa H, Kotani T, Ohtaki S, Nakamura M, Yamazaki I

出版信息

Biochem Biophys Res Commun. 1985 Feb 28;127(1):8-14. doi: 10.1016/s0006-291x(85)80118-2.

Abstract

A rapid method was developed for purification of hog thyroid peroxidase by immunoaffinity chromatography on a column of Sepharose 4B coupled to a monoclonal antibody to the peroxidase. The purified enzyme had a specific activity of 194 units/mg and showed the same absorption spectrum in the Soret and visible regions as that of the enzyme purified after trypsin treatment. The ratio of A413 nm to A280 nm was 0.24, being much less than that for the trypsinized enzymes. Upon sodium dodecyl sulfate-polyacrylamide gel electrophoresis, it gave a broad protein band in the 100,000-dalton region. It is concluded that the preparation purified in this study represents a native form of thyroid peroxidase.

摘要

开发了一种快速方法，通过在与抗过氧化物酶单克隆抗体偶联的琼脂糖4B柱上进行免疫亲和色谱法来纯化猪甲状腺过氧化物酶。纯化后的酶比活性为194单位/毫克，在Soret和可见光区域的吸收光谱与胰蛋白酶处理后纯化的酶相同。A413nm与A280nm的比值为0.24，远低于经胰蛋白酶处理的酶。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中，它在100,000道尔顿区域给出一条宽蛋白带。得出的结论是，本研究中纯化的制剂代表甲状腺过氧化物酶的天然形式。