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A Biotin Ligase-Based Assay for the Quantification of the Cytosolic Delivery of Therapeutic Proteins


Verdurmen, Wouter P R; Mazlami, Marigona; Plückthun, Andreas (2017). A Biotin Ligase-Based Assay for the Quantification of the Cytosolic Delivery of Therapeutic Proteins. Methods in Molecular Biology, 1575:223-236.

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.

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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Additional indexing

Item Type:Journal Article, refereed, original work
Communities & Collections:04 Faculty of Medicine > Department of Biochemistry
07 Faculty of Science > Department of Biochemistry
Dewey Decimal Classification:570 Life sciences; biology
610 Medicine & health
Language:English
Date:2017
Deposited On:12 Apr 2017 12:55
Last Modified:19 Feb 2018 07:50
Publisher:Springer
ISSN:1064-3745
OA Status:Closed
Publisher DOI:https://doi.org/10.1007/978-1-4939-6857-2_14
PubMed ID:28255884

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