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Assessment of the effect of triton X‐114 on the physicochemical properties of an antibody fragment
Authors:Luciana P Malpiedi  Bibiana B Nerli  Dulcineia S P Abdalla  Adalberto Pessoa Jr
Institution:1. Biochemical and Pharmaceutical Technology Dept./FCF, University of S?o Paulo. Av. Prof. Lineu Prestes, S?o Paulo, Brazil;2. Physical Chemistry Dept., Faculty of Biochemistry and Pharmaceutical Sciences, National University of Rosario, Rosario, Argentina;3. Dept. of Clinical Analysis and Toxicology, University of S?o Paulo, S?o Paulo, Brazil
Abstract:The effect of Triton X‐114 on the physicochemical properties of a single‐chain antibody fragment (scFv) has been studied. According to the far UV circular dichroism spectroscopy, the secondary structure of the recombinant antibody was not significantly affected by the presence of Triton. From the antibody tertiary structure analysis, it was found that the surfactant could be located around the tryptophan molecules accessible to the solvent, diminishing the polarity of its environment but maintaining most of the protein structure integrity. However, in certain conditions of high temperature and high concentration of denaturant molecules, the presence of TX could compromise the antibody fragment stability. These results represent a previous step in designing scFv purification protocols and should be considered prior to developing scFv liquid–liquid extraction procedures. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 30:554–561, 2014
Keywords:nonionic surfactant  single‐chain antibody fragment  spectroscopic study
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