喷雾干燥氧化锆微球在从细胞培养上清液中分离免疫球蛋白中的应用。

Use of spray-dried zirconia microspheres in the separation of immunoglobulins from cell culture supernatant.

作者信息

Subramanian A, Carr P W, McNeff C V

机构信息

University of Minnesota, Department of Biosystems and Agricultural Engineering, St. Paul 55108, USA.

出版信息

J Chromatogr A. 2000 Aug 18;890(1):15-23. doi: 10.1016/s0021-9673(00)00289-2.

DOI:10.1016/s0021-9673(00)00289-2

PMID:10976790

Abstract

A method suitable for the isolation of monoclonal antibodies (MAbs) on novel zirconia microspheres (20-30 microm) is described. Zirconia microspheres were generated by spray drying colloidal zirconia. Spray-dried zirconia microspheres were further classified and characterized by X-ray diffraction, BET porosimetry and scanning electron microscopy. Spray-dried zirconia microspheres were modified with ethylenediamine-N,N'-tetra(methylenephosphonic) acid (EDTPA) to create a cation-exchange chromatographic support. The chromatographic behavior of a semi-preparative column packed with EDTPA-modified zirconia microspheres was evaluated and implications for scale-up are provided. EDTPA-modified zirconia microspheres were further used to purify MAbs from cell culture supernatant. Analysis by enzyme linked immunosorbent assay and gel electrophoresis demonstrate that MAbs can be recovered from a cell culture supernatant at high yield (92-98%) and high purity (>95%) in a single chromatographic step.

摘要

本文描述了一种适用于在新型氧化锆微球（20 - 30微米）上分离单克隆抗体（MAbs）的方法。氧化锆微球通过喷雾干燥胶体氧化锆生成。喷雾干燥的氧化锆微球通过X射线衍射、BET孔隙率测定法和扫描电子显微镜进一步分类和表征。喷雾干燥的氧化锆微球用乙二胺 - N,N'-四（亚甲基膦酸）（EDTPA）进行修饰，以创建阳离子交换色谱支持物。评估了填充有EDTPA修饰的氧化锆微球的半制备柱的色谱行为，并提供了放大的相关信息。EDTPA修饰的氧化锆微球进一步用于从细胞培养上清液中纯化单克隆抗体。通过酶联免疫吸附测定和凝胶电泳分析表明，单克隆抗体可以在单个色谱步骤中以高产率（92 - 98%）和高纯度（>95%）从细胞培养上清液中回收。