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pH-shift stress on antibodies
Hiroshi Imamura1, Shinya Honda2
1Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, Tsukuba, Japan; Department of Applied Chemistry, College of Life Sciences, Ritsumeikan University, Kusatsu, Japan.
Antibody unfolding and aggregation have been gaining increasing concern as the need for pharmaceutical antibodies with high quality has increased. Assays for continuous application of severe stress to antibodies, which are typically employed to evaluate tolerance to aggregation or to understand the mechanism of aggregation, induce both unfolding and aggregation simultaneously. In this chapter, we describe the pH-shift stress on antibodies that uses a low pH solution to unfold the antibody and a neutralizing solution to induce folding and the aggregation. This pH-shift technique with light scattering analysis will be a useful assay to evaluate antibody aggregation at a neutral pH and ambient temperature without generation of unfolded molecules during aggregation.
Antibody unfolding and aggregation have been gaining increasing concern as the need for pharmaceutical antibodies with high quality has increased. Assays for continuous application of severe stress to antibodies, which are typically employed to evaluate tolerance to aggregation or to understand the mechanism of aggregation, induce both unfolding and aggregation simultaneously. In this chapter, we describe the pH-shift stress on antibodies that uses a low pH solution to unfold the antibody and a neutralizing solution to induce folding and the aggregation. This pH-shift technique with light scattering analysis will be a useful assay to evaluate antibody aggregation at a neutral pH and ambient temperature without generation of unfolded molecules during aggregation.
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