Banca de DEFESA: ANA LAURA OLIVEIRA DE SÁ LEITÃO

Uma banca de DEFESA de MESTRADO foi cadastrada pelo programa.
DISCENTE : ANA LAURA OLIVEIRA DE SÁ LEITÃO
DATA : 08/02/2017
HORA: 14:30
LOCAL: AUDITÓRIO DO PPGEQ
TÍTULO:

"Evaluation of cell disruption methods and of different metals immobilized on Streamline Chelating resin for purification 503 antigen of  Leishmania  i. Chagasi."

PALAVRAS-CHAVES:

Expanded bed adsorption, purification, metal ion, 503 antigen.

PÁGINAS: 96
GRANDE ÁREA: Engenharias
ÁREA: Engenharia Química
RESUMO:

The growing interest of the bioprocess industry for the purified recombinant antigens is mainly due they can be used as a tool for the vaccines production. However, the application of these antigens requires a quite higher degree of purity thus techniques that could reduce the downstream processing cost it would be important. The Expanded Bed Adsorption (EBA) has been shown as a good alternative since it is simple, and combines the clarification, concentration and purification at only step. This study aims investigating the recovery and purification of the 503 antigen from Leishmania i. chagasi expressed in E. coli as well as the simultaneous removal of the endotoxin using EBA coupled to IMAC. First, the best method for the cellular disruption for obtaining the higher antigen content was assayed. The methods studied used were assayed by means of factoral planning using Lysozyme, beads, urea as well as EDTA. Next, adsorption tests in batch were used in order to investigate the use of five metal ions (Cu²⁺, Ni²⁺, Zn²⁺, Co²⁺, Fe³⁺) at concentration of 0.1 M, 0.5 M and 0.8 M coupled to the Streamline chelating resin in the adsorption capacity of the 503 antigen. The metal showing the best adsorption capacity was chosen for the EBA experiments. EBA experiments were carried out using a column of 2.6 cm in diameter (30 cm height), coupled to a peristaltic pump. Results showed that higher quantity for the total protein as well as the 503 antigen was obtained ureia at 8.0 M. Also, the best batch adsorption was shown by the Cu²⁺ metal showing 503 antigen adsorption capacity of 0.102; 0.128 and 0.111 mg/mL of adsorbent for concentrations of 0.1, 0.5 and 0.8 M, respectively. Also, EBA protocol using Streamline chelating coupled copper eluted witht 0.6 M of imidazole was able to recovery only 4.0 % 503 antigen.

 

 

MEMBROS DA BANCA:
Presidente - 1346198 - EVERALDO SILVINO DOS SANTOS
Interno - 347401 - GORETE RIBEIRO DE MACEDO
Externo à Instituição - FRANCISCO CANINDE DE SOUSA JUNIOR - UFRN
Externo à Instituição - IGOR TADEU LAZZAROTTO BRESOLIN - UNIFESP