Efficient wheat germ agglutinin purification with a chitosan-based affinity chromatographic matrix

✍ Scribed by María F. Baieli; Nicolás Urtasun; María V. Miranda; Osvaldo Cascone; Federico J. Wolman

• Publisher: John Wiley and Sons
• Year: 2012
• Tongue: English
• Weight: 291 KB
• Volume: 35
• Category: Article
• ISSN: 1615-9306
• DOI: 10.1002/jssc.201100730

✦ Synopsis

Abstract

An efficient affinity chromatographic matrix based on chitosan for wheat germ agglutinin (WGA) purification was developed. The matrices assayed consisted of chitosan mini‐spheres cross‐linked with epichlorhydrin 45, 250 or 500 mM. The maximum adsorption capacity of pure WGA – calculated from the corresponding isotherms – was between 43.2 and 48.9 mg/g at pH 5.0 and between 16.6 and 27.6 mg/g at pH 8.5. However, the adsorption of agglutinin from wheat germ extract was higher at pH 8.5. In addition, 0.5 g of mini‐spheres cross‐linked with epichlorhydrin 250 mM adsorbed 94.5% of the WGA present in 5 mL of the concentrated extract. Acetic acid was able to elute 100% of the adsorbed WGA. The purity of the WGA obtained was greater than 95% and the purification factor was 56.8. The matrix was able to maintain an efficient performance of the purification process for three consecutive cycles. A new method to monitor the purification process by RP‐HPLC was developed.