使用高效液相色谱法从腹水液中快速高效地纯化小鼠单克隆抗体。

Rapid and efficient purification of mouse monoclonal antibodies from ascites fluid using high performance liquid chromatography.

作者信息

Burchiel S W, Billman J R, Alber T R

出版信息

J Immunol Methods. 1984 Apr 13;69(1):33-42. doi: 10.1016/0022-1759(84)90274-6.

Abstract

Techniques for the rapid and efficient purification of mouse monoclonal antibodies from murine ascites are described that utilize anion exchange and gel permeation chromatography using high performance liquid chromatography (HPLC). Anion exchange chromatography was performed at neutral pH using a hydrophilic resin conjugated with a substituted amine (Mono Q column, Pharmacia Fine Chemicals). Various subclasses of mouse IgG monoclonals were assessed for their binding to this matrix, with all of the antibodies tested eluting at relatively low concentrations of sodium chloride. In some instances, a protein tentatively identified as transferrin was co-purified using this anion exchange procedure. However, this protein was easily removed from the IgG using gel permeation chromatography (Bio-Sil TSK-250, Bio-Rad), also performed at neutral pH.

摘要

本文描述了利用阴离子交换和凝胶渗透色谱法结合高效液相色谱（HPLC）从鼠腹水快速高效纯化小鼠单克隆抗体的技术。阴离子交换色谱在中性pH下使用与取代胺偶联的亲水性树脂（Mono Q柱，Pharmacia Fine Chemicals）进行。评估了小鼠IgG单克隆抗体的各种亚类与该基质的结合情况，所有测试的抗体均在相对低浓度的氯化钠下洗脱。在某些情况下，使用这种阴离子交换方法共纯化了一种暂定为转铁蛋白的蛋白质。然而，使用同样在中性pH下进行的凝胶渗透色谱法（Bio-Sil TSK-250，Bio-Rad）可轻松从IgG中去除该蛋白质。