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Efficient wheat germ agglutinin purification with a chitosan‐based affinity chromatographic matrix
Author(s) -
Baieli María F.,
Urtasun Nicolás,
Miranda María V.,
Cascone Osvaldo,
Wolman Federico J.
Publication year - 2012
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201100730
Subject(s) - wheat germ agglutinin , chromatography , chemistry , adsorption , chitosan , matrix (chemical analysis) , elution , agglutinin , affinity chromatography , germ , acetic acid , lectin , biochemistry , organic chemistry , enzyme , biology , microbiology and biotechnology
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.