Extraction of Human IgG in Thermo-Responsive Aqueous Two-Phase Systems: Assessment of Structural Stability by Circular Dichroism

被引:4
|
作者
Borlido, Luis [1 ]
Azevedo, Ana M. [1 ]
Raquel Aires-Barros, M. [1 ]
机构
[1] Inst Super Tecn, Ctr Biol & Chem Engn, IBB, P-1049001 Lisbon, Portugal
关键词
aqueous two-phase system; circular dichroism; immunoglobulin G; temperature sensitive polymers; CONFORMATION; ANTIBODIES; IMMUNOGLOBULIN; AGGREGATION; PROTEIN;
D O I
10.1080/01496395.2010.507441
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
The structural stability of human IgG was studied throughout the extraction process with thermo-responsive UCON/dextran aqueous two-phase systems. Systems composed by 8% UCON and 5% dextran were assessed in both acetate pH 5 and phosphate pH 7. The extraction of IgG to the top phase was found to be favored at pH 5 reaching 85% yield with UCON 50HB-3520. During the extraction process conducted at 25 degrees C no alteration in the secondary structure, evaluated by circular dichroism, was observed. The back-extraction step was induced by the thermo-precipitation of the UCON polymers at 59 degrees C. At both pH values, mild changes in the secondary structure of IgG were observed for the 50HB-3520 and 50HB-5100 polymers while 50HB-2000 showed an increase in -sheet content. The overall yield of the process was as high as 82% determined for the UCON 50HB-3520 system at pH 5.
引用
收藏
页码:2171 / 2179
页数:9
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