生物-曼吉诺斯结合疫苗中载体蛋白分子表征的物理化学方法

Anais do IV International Symposium on Immunobiological e VII Seminário Anual Científico e Tecnológico de Bio-Manguinhos Pub Date : 1900-01-01 DOI:10.35259/isi.sact.2019_32537

I. Corrêa, R. Bastos, P. Jurgilas, H. Nascimento

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引用次数: 0

摘要

方法:使用三种不同的色谱柱(Zorbax®GF450, TSK®G4000 SWXL和SuperdexTM 200)对两种不同的TT样品(TT1和TT2)进行SEC色谱分析。采用SDS-PAGE、IEF-PAGE和NATIVE-PAGE等电泳技术评价电泳谱。在温度25℃-85℃范围内，分别用圆二色性和荧光光谱分析了二级和三级结构的稳定性。所有蛋白均经过胰蛋白酶水解，目的是通过RPC色谱获得肽图。所有分析均以TT1为标准。

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Physical-chemical methodologies for molecular characterization of carrier proteins used in Bio-Manguinhos conjugated vaccines

Methodology: SEC chromatographic profile of two different samples of TT (TT1 and TT2) were performed using three different columns (Zorbax®GF450, TSK®G4000 SWXL and SuperdexTM 200). Electrophoretic techniques such as SDS-PAGE, IEF-PAGE and NATIVE-PAGE were used to evaluate electrophoretic profile. Stability of secondary and tertiary structures were analyzed by circular dichroism and fluorescence spectroscopy respectively, with temperature varying between 25°C-85°C. All proteins were subjected to tryptic hydrolysis aiming obtain a peptide map by RPC chromatography. TT1 was used as standard for all analysis.

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Anais do IV International Symposium on Immunobiological e VII Seminário Anual Científico e Tecnológico de Bio-Manguinhos

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