LECTINA LIGADORA DA MANOSE: PROPRIEDADES, ISOLAMENTO E PURIFICAÇÃO

The mannose-binding lectin (MBL) is a human serum protein, which is secreted by the liver and is calcium-dependent. The MBL is an important constituent of the innate immune system and it may act by activating the complement system through the lectins pathway and by its characteristic of being an opsonin, facilitating the pathogens phagocytosis. This protein is associated to infectious and autoimmune disease, interfering in the individual susceptibility and in the disease severity. The goal of this work was isolating and purifying the MBL from human serum, through a chromatographic method, in order to use it in a future monoclonal antibody production. The method used to isolate and purify the MBL was the affinity chromatography, performed in columns. The method’s protocol was developed during the study, based on information from previous published reports. In this study, some columns of different sizes were used and the fixed phase was prepared with Sepharose 6B-D-mannose. The fractions obtained through the chromatography were taken to spectrophotometer analysis, using the absorbance as the determination criteria for the MBL presence in the samples. The protein solution obtained through the chromatography method was concentrated to allow the next step, the eletrophoresis identification. The identification result indicated that the isolated protein is the MBL. The chromatographic method developed during this study showed to be able to isolate and purify the MBL; nevertheless, some adjusts still need to be made to improve the method, in order to enhance its speedy and efficiency