Analysis of proteins by agarose native gel electrophoresis in the presence of solvent additives

Solvent additives, including NaCl, arginine hydrochloride (ArgHCl), glycine and sucrose, are used to enhance protein stability or reduce protein aggregation. Here, we studied the effects of these additives on proteins using agarose native gel electrophoresis. Since these additives are used at relati...

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Bibliographic Details
Published in:International journal of biological macromolecules 2022-02, Vol.198, p.26-36
Main Authors: Tomioka, Yui, Arakawa, Tsutomu, Akuta, Teruo, Nakagawa, Masataka, Ishibashi, Matsujiro
Format: Article
Language:English
Subjects:
Agarose native gel
Arginine
Muramidase
Soluble aggregates
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Summary:Solvent additives, including NaCl, arginine hydrochloride (ArgHCl), glycine and sucrose, are used to enhance protein stability or reduce protein aggregation. Here, we studied the effects of these additives on proteins using agarose native gel electrophoresis. Since these additives are used at relatively high concentration, we first confirmed that they do not interfere with the performance of the native gel electrophoresis. Agarose native gel electrophoresis showed that aggregation of bovine serum albumin (BSA) induced by heating was slightly reduced by NaCl and ArgHCl. On the contrary, glycine and sucrose had marginal effects. ArgHCl and NaCl promoted heat aggregation of monoclonal antibody (mAb), while glycine and sucrose stabilized the native mAb. Arginine methyl ester inhibited heat aggregation of lysozyme and, to a much lesser extent, BSA. These results show that agarose native gel electrophoresis can be used to analyze the effects of solvent additives on proteins subjected to heat stresses. SYPRO Orange that stains only unfolded proteins confirmed unfolded structures of soluble aggregates.
ISSN:0141-8130
1879-0003
1879-0003
DOI:10.1016/j.ijbiomac.2021.12.084