Isolamento de glicoproteínas através de cromatografia de afinidade em colunas contendo lectinas imobilizadas

by dos Santos, Teodomiro Gomes

Bauhinia monandra leaf lectin (BmoLL) and the isolectins from Cratylia mollis seeds (Cra Iso 1,2,3) isolated in the Laboratory of Glycoproteins from UFPE were immobilized in polyvinyl alcohol-glutaraldehyde (PVAglurataraldehyde)and Sepharose-4B, respectively. The matrices Cra Iso 1,2,3-Sepharose, SpiL-Sepharose (seed lectin from Swartzia pickellii immobilized in Sepharose-4B), and BmoLL-PVA-glutaraldehyde were evaluated in relation tothe binding efficiency of ovalbumin and isolation of glycoproteins from fetal bovine serum and human colostrum. The yields of immobilization to Cra Iso 1,2,3-Sepharose and BmoLL-PVA-glutaraldehyde were 90 % and 50%, respectively. Distinct experimental conditions using the matrices were evaluated (column chromatography and batch assay) which gave different results. Proteins with electrophoretic migration similar to secretory IgA from human colostrum and similar to fetuin were obtained after chromatography of humancolostrum in BmoLL-PVA-glutaraldehyde column and fetal bovine serum in Cra Iso 1,2,3-Sepharose column, respectively. Ovalbumin was retained in Cra Iso 1,2,3-Sepharose using batch assays. The matrices were stable under the used chromatographic conditions. Cra Iso 1,2,3-Sepharose and BmoLL-PVAglutaraldehyde can be included in the group of matrices to affinity chromatography with the aim to obtain glycoconjugates. SpiL-Sepharose wasnot efficient to bind ovalbumin or fetal bovine serum glycoproteins, under the evaluated experimental conditions