Abstract
High pressure homogenization (HPH) is a commonly used method for cell lysis of Escherichia coli in order to release intracellularly produced recombinant proteins. For misfolded proteins in E. coli, focus is often put on the development of a suitable solubilization and refolding protocol. However, HPH can be a critical unit operation influencing inclusion body (IB) quality and, subsequently, refolding yields. Here, a protocol for homogenization and IB washing is presented in combination with analytical methods suitable to evaluate these unit operations. The protocol is based on a multivariate approach to identify suitable conditions during HPH. Furthermore, the described workflow is easily scalable and can, therefore, also be used if fixed homogenization conditions are already established.
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Ebner, J., Sedlmayr, V., Klausser, R. (2023). High Pressure Homogenization for Inclusion Body Isolation. In: Kopp, J., Spadiut, O. (eds) Inclusion Bodies. Methods in Molecular Biology, vol 2617. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2930-7_9
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DOI: https://doi.org/10.1007/978-1-0716-2930-7_9
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Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-2929-1
Online ISBN: 978-1-0716-2930-7
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