Purification of secreted α-amylases by immunoaffinity chromatography with cross-reactive antibody
Purification of secreted α-amylases by immunoaffinity chromatography with cross-reactive antibody
复制标题
使用交叉反应抗体通过免疫亲和层析纯化分泌的 α-淀粉酶
DOI：
10.1007/bf00170221
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发表时间：
1994
影响因子：
5
通讯作者：
M. Terashima
中科院分区：
文献类型：
--
作者：
S. Katoh;M. Terashima
利用免疫亲和层析技术，利用交叉反应抗体对重组酵母表达和分泌的水稻α-淀粉酶的两种同工酶进行了纯化。针对部分纯化的大麦α-淀粉酶的抗体通过交叉反应吸附发酵液中的大米α-淀粉酶。以这些抗体为配体，采用一步免疫亲和层析法对水稻α-淀粉酶进行了高度浓缩和纯化。由于来自大麦芽的大麦α-淀粉酶（抗原）和来自酵母分泌的大米α-淀粉酶（靶蛋白）之间的污染杂质的差异，获得了洗脱的高纯度α-淀粉酶，而无需使用高度纯化的抗原用于免疫。在免疫亲和层析中使用交叉反应性抗体可用于在缺乏足够量和足够高纯度的待纯化靶蛋白的情况下纯化重组蛋白。
Two isozymes of rice α-amylases expressed and secreted by recombinant yeast were purified by immunoaffinity chromatography by using cross-reactive antibody. Antibodies raised against partially purified barley α-amylase adsorbed rice α-amylases in fermentation broth by a cross-reaction. By use of these antibodies as ligands, rice α-amylases were concentrated and purified to a high degree in one-step immunoaffinity chromatography. Because of the differences in the contaminating impurities between the barley α-amylase (antigen) from barley malt and rice α-amylases (target protein) secreted from yeast, the high purity of eluted α-amylases was attained without the use of highly purified antigen for immunization. Utilization of cross-reactive antibodies in immunoaffinity chromatography is useful for the purification of recombinant proteins in the absence of a sufficient amount and high enough purity of the target proteins to be purified.