Abstract
The efficient delivery of external proteins from the external milieu to the cytosol of mammalian cells has great potential for both scientific investigations and future therapies. However, when assessing the cellular uptake of proteins, it is often difficult to distinguish between proteins that are stuck in the endosomes and those that have escaped into the cytosol. Here, we describe a method employing the prokaryotic enzyme biotin ligase that overcomes this problem and which can be employed for a highly sensitive quantification of cytosolic protein delivery.
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Acknowledgments
The research leading to these methods has received support from the EU/EFPIA Innovative Medicines Initiative Joint Undertaking COMPACT grant no 115363. W.P.R.V. was supported by a Rubicon grant (Project number: 825.12.002) from the Netherlands Organisation for Scientific Research and the Forschungskredit from the University of Zurich.
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Verdurmen, W.P.R., Mazlami, M., Plückthun, A. (2017). A Biotin Ligase-Based Assay for the Quantification of the Cytosolic Delivery of Therapeutic Proteins. In: Tiller, T. (eds) Synthetic Antibodies. Methods in Molecular Biology, vol 1575. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6857-2_14
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DOI: https://doi.org/10.1007/978-1-4939-6857-2_14
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