从小鼠肝脏中通过抗体亲和纯化新型α-L-岩藻糖苷酶。

Antibody-affinity purification of novel alpha-L-fucosidase from mouse liver.

作者信息

Laury-Kleintop L D, Damjanov I, Alhadeff J A

机构信息

Department of Chemistry, Lehigh University, Bethlehem, PA 18015.

出版信息

Biochem J. 1987 Jul 15;245(2):589-93. doi: 10.1042/bj2450589.

DOI:10.1042/bj2450589

PMID:3663178

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1148162/

Abstract

Previous studies have documented the presence of a novel alpha-L-fucosidase in mouse liver that contains unique basic isoelectric forms and that is antigenically similar to, but not identical with, human liver alpha-L-fucosidase [Laury-Kleintop, Damjanov & Alhadeff (1985) Biochem. J. 230, 75-82]. In the present investigation, mouse liver alpha-L-fucosidase was purified approx. 26,500-fold in 10% overall yield by antibody-affinity chromatography with the IgG fraction of goat anti-(human alpha-L-fucosidase) antibody coupled to Sepharose 4B. Native polyacrylamide-gel electrophoresis and SDS/polyacrylamide-gel electrophoresis indicated that the mouse fucosidase is highly purified if not homogeneous. Isoelectric focusing demonstrated that all enzymic forms found in crude mouse liver supernatant fluids were purified by the antibody-affinity procedure.

摘要

先前的研究已证明，小鼠肝脏中存在一种新型α-L-岩藻糖苷酶，它具有独特的碱性等电形式，在抗原性上与人类肝脏α-L-岩藻糖苷酶相似，但并不相同[劳里-克莱因托普、达米亚诺夫和阿尔哈杰夫（1985年）《生物化学杂志》230卷，75 - 82页]。在本研究中，通过将山羊抗（人类α-L-岩藻糖苷酶）抗体的IgG组分偶联到琼脂糖4B上进行抗体亲和层析，小鼠肝脏α-L-岩藻糖苷酶被纯化了约26,500倍，总产率为10%。天然聚丙烯酰胺凝胶电泳和SDS/聚丙烯酰胺凝胶电泳表明，即使不是纯一的，小鼠岩藻糖苷酶也已高度纯化。等电聚焦显示，粗制小鼠肝脏上清液中发现的所有酶形式都通过抗体亲和程序得到了纯化。