Chitosan microparticle-based immunoaffinity chromatography supports prepared by membrane emulsification technique: Characterization and application.

Abstract

In this study, chitosan microparticles intended to be used as matrix support for immunoaffinity chromatography were prepared by membrane emulsification technique. Regular spherical chitosan microparticles were obtained with an average diameter of 62.9μm and a size distribution index of 1.9 and showed stable under wide pH range (4.0 to 10.0) and various chemical conditions. Size distribution and chemical stability of the prepared chitosan microparticles were relatively close to those of Sepharose 4B. Polyclonal antibody against zearalenone (ZEN) as a model ligand was coupled onto chitosan microparticles or Sepharose 4B to obtain immunoaffinity columns. Further characterization indicated that chitosan microparticle-based column exhibited a maximum binding capacity of 6.1μg/mL gel, higher than that (3.1μg/mL gel) of Sepharose 4B-based column, and a column-to-column variation of 6.0%, slightly lower than that (8.6%) of Sepharose 4B-based column. When challenged with cornmeal samples fortified with ZEN at 50 and 100ng/g, satisfied recoveries of 92.1-100.2% with relative standard deviation (RSD)≤7.6% and 88.3-107.3% with RSD≤7.6% were both yielded by chitosan microparticle- and Sepharose 4B-based column, respectively, indicating that the obtained chitosan microparticles could be hopefully used as matrix support for immunoaffinity chromatography with reproducible extraction behavior and high extraction efficiency.