Analytical Sciences

Rapid Purification of Immunoglobulin G Using a Protein A-​immobilized Monolithic Spin Column with Hydrophilic Polymers

Shigenori OTA,* Yuko YUI,** Tsutomu SATO,** Noriko YOSHIMOTO,* and Shuichi YAMAMOTO*

*Bio-Process Engineering Laboratory, Graduate School of Yamaguchi University Biomedical Engineering Center (YUBEC) 2-16-1 Tokiwadai, Ube 755-8611, Japan
**GL Sciences Inc., 237-2 Sayamagahara, Iruma, Saitama 358-0032, Japan

A rapid purification method was developed for antibody production in Chinese hamster ovary (CHO) cells using a Protein A-immobilized monolithic silica spin column with hydrophilic polymers. Monolithic silica modified with copolymers of 2-hydroxyethylmethacrylate (HEMA) and glycidyl methacrylate (GMA) showed lower non-specific protein absorption than that modified with a silane reagent. The epoxy group of GMA was converted to an amino group, and Protein A was modified by the coupling reagent. The amount of immobilized Protein A was controlled by changing the ratio of GMA to HEMA and the mesopore size of monolith. A modified monolith disk was fixed to a spin column for rapid antibody purification. The linear curves (for the antibody concentrations over 10 – 300 μg/mL) had a correlation coefficient of >0.999. Our column had various analytical advantages over previously reported columns, including a shorter preparation time (<10 min) and smaller sample volumes for purification with Protein A-immobilized agarose.

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