title: Affinity matrices of Cratylia mollis seed lectins for isolation of glycoproteins from complex protein mixtures.
abstract: This work reports the use of matrices containing Cratylia mollis lectins (Cramoll 1,2,3-Sepharose and Cramoll 3-Sepharose) for isolation of glycoproteins from fetal bovine serum, human colostrum, hen egg white, and human blood plasma. Cramoll 1,2,3-Sepharose was able to bind a glycoprotein from fetal bovine serum which showed the same fetuin electrophoretic profile. The data indicate that this protein adsorbed to the matrix by interaction with Cramoll 3. Cramoll 1,2,3-Sepharose was not efficient to retain glycoproteins from human colostrum or commercial ovalbumin. Cramoll 3-Sepharose bound ovalbumin, and the support retained protein from hen egg white. Protein peaks eluted from the column with 1.0 M NaCl or 0.3 M galactose showed apparent molecular mass of ovalbumin. Two main proteins from blood plasma with apparent molecular mass 67 (similar to albumin) and 50 kDa (similar to fetuin) adsorbed on Cramoll 3-Sepharose and were eluted with 1.0 M NaCl as a single peak. Elution of adsorbed plasma proteins with 0.3 M galactose was less selective than with 1.0 M NaCl as revealed by SDS-PAGE. In conclusion, the Cramoll 1,2,3-Sepharose and Cramoll 3-Sepharose matrices were useful to separate glycoproteins from complex protein mixtures, and the adsorption phenomena was a carbohydrate-dependent event.
authors.affiliation: Departamento de Bioquímica, Centro de Ciências Biológicas, Universidade Federal de Pernambuco, Recife, Pernambuco, 50670-420, Brazil.
authors.forename: Thiago Henrique
authors.initials: TH
authors.lastname: Napoleão
authors.forename: Teodomiro Gomes
authors.initials: TG
authors.lastname: dos Santos-Filho
authors.forename: Emmanuel Viana
authors.initials: EV
authors.lastname: Pontual
authors.forename: Rodrigo da Silva
authors.initials: Rda S
authors.lastname: Ferreira
authors.forename: Luana Cassandra Breitenbach Barroso
authors.initials: LC
authors.lastname: Coelho
authors.forename: Patrícia Maria Guedes
authors.initials: PM
authors.lastname: Paiva
mesh_terms: D000818:Animals; D002417:Cattle; D002846:Chromatography, Affinity / Q000379:methods*; D007887:Fabaceae / Q000737:chemistry; D006023:Glycoproteins / Q000302:isolation & purification*; D006801:Humans; D037102:Lectins / Q000737:chemistry*; D010047:Ovalbumin / Q000302:isolation & purification; D012639:Seeds / Q000737:chemistry; D012685:Sepharose
chemical_list: D006023:Glycoproteins; D037102:Lectins; D010047:Ovalbumin; D012685:Sepharose