Affinity Matrices of Cratylia mollis Seed Lectins for Isolation of Glycoproteins from Complex Protein Mixtures

作者：Napoleao Thiago Henrique; dos Santos Filho Teodomiro Gomes; Pontual Emmanuel Viana; Ferreira Rodrigo da Silva; Breitenbach Barroso Coelho Luana Cassandra; Guedes Paiva Patricia Maria

来源：Applied Biochemistry and Biotechnology, 2013, 171(3): 744-755.

DOI：10.1007/s12010-013-0403-z

• Cratylia mollis
• Lectin affinity chromatography
• Ovalbumin
• Fetuin
• Albumin

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.

• 出版日期2013-10