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

Autor: Baieli MF; School of Pharmacy and Biochemistry, University of Buenos Aires, Junín 956, Buenos Aires, Argentina., Urtasun N, Miranda MV, Cascone O, Wolman FJ
Jazyk: angličtina
Zdroj: Journal of separation science [J Sep Sci] 2012 Jan; Vol. 35 (2), pp. 231-8.
DOI: 10.1002/jssc.201100730
Abstrakt: 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.
(Copyright © 2012 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim.)
Databáze: MEDLINE
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