PURIFICATION AND OPTIMIZATION OF MANNOSE BINDING LECTIN FROM BOVINE’S LUNG TISSUE

Abstract

Mannose binding lectin (MBL) is calcium dependent serum protein that used as a natural immune response and made in the liver. MBL works by binding to carbohydrates on the surface of pathogens where it can activate the complement system directly or act as opsonin. The MBL deficiency due to mutation of MBL2 gene cause people with this mutation has high risk of infection. Low MBL serum levels will enhance risk of infections whereas high MBL serum levels will increase inflammatory diseases, transplant rejection and diabetic nephropathy due to its immune system property. This study suggested an effective purification method to purify MBL for human based on bovine lung tissue since availability of human sample was limited. There were two types of column used for purification of MBL from bovine’s lung tissue in this study which is GlcNAc and mannose-Sepharose column based on previous study. The protein concentration detect in sample using Bradford assay show that mannose-Sepharose column can purified MBL better than GlcNAc column. This study proceeds with SDS-PAGE to prove the present of MBL protein in bovine’s lung tissue. However there was no band appeared on the gel after electrophoresis. The factors predicted can caused this mistake were happen during sample preparation, column preparation, present of detergent, coomassie dye binding used and method apply during SDS-PAGE.