INVESTIGADORES
MIRANDA Maria Victoria
artículos
Título:
Efficient wheat germ agglutinin purification with a chitosan affinity chromatographic matrix.
Autor/es:
BAIELI FERNANDA; URTASUN NICOLÁS; MARIA V MIRANDA; CASCONE O; FEDERICO WOLMAN
Revista:
JOURNAL OF SEPARATION SCIENCE - (Online)
Editorial:
Wiley
Referencias:
Año: 2012 vol. 35 p. 231 - 238
ISSN:
1615-9314
Resumen:
An efficient affinity chromatographic matrix based on chitosan for wheat germ agglutinin
(WGA) purification was developed. The matrices assayed consisted of chitosan minispheres
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 250mM adsorbed 94.5% of the WGA
present in 5mL 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.