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Please use this identifier to cite or link to this item: http://hdl.handle.net/10119/15877

Title: Enhanced Adsorption of a Protein-nanocarrier Complex onto Cell Membranes through a High Freeze Concentration by a Polyampholyte Cryoprotectant
Authors: Ahmed, Sana
Miyawaki, Osato
Matsumura, Kazuaki
Keywords: cytoplasmic delivery
freeze concentration
drug delivery
Issue Date: 2018-01-23
Publisher: American Chemical Society
Magazine name: Langmuir
Volume: 34
Number: 6
Start page: 2352
End page: 2362
DOI: 10.1021/acs.langmuir.7b03622
Abstract: Transportation of biomolecules into cells is of great importance in tissue engineering and as stimulation for antitumor immune cells. Previous freezing strategies at ultra-cold temperatures (−80 °C) used for intracellular transportation exhibit certain limitations such as extended time requirements and harsh delivery system conditions. Thus, the need remains to develop simplified methods for safe nanomaterial delivery. Here, we demonstrated a unique strategy based on ice crystallization-induced freeze concentration for protein intracellular delivery in combination with a polyampholyte cryoprotectant. We found that upon sustained lowering of temperatures from −6 to −20 °C over a short duration, the adsorption of proteins onto the peripheral cell membrane was markedly increased through facile ice crystallization-induced freeze concentration. Furthermore, we proposed a “freeze concentration factor” (α) that depends on the freezing point depression and is estimated from analysis of the fraction of frozen water. Notably, the α values of polyampholyte cryoprotectant were 6-fold higher than those of the currently used cryoprotectant dimethyl sulfoxide (DMSO) at particular temperatures of interest. Our results illustrate that the presence of a polyampholyte cryoprotectant significantly enhanced adsorption of the protein/nanocarrier complex onto membranes compared with that obtained with DMSO because of high freeze concentration. The present study demonstrated the direct relationship of freezing with penetration of proteins across the periphery of the cell membrane by means of increased concentration during freezing. These results may be useful in providing a guideline for the intracellular delivery of biomacromolecules using ice crystallization-induced continuous freezing combined with polyampholyte cryoprotectants.
Rights: Sana Ahmed, Osato Miyawaki, Kazuaki Matsumura, Langmuir, 2018, 34(6), pp.2352-2362. This document is the Accepted Manuscript version of a Published Work that appeared in final form in Langmuir, copyright (c) American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see http://dx.doi.org/10.1021/acs.langmuir.7b03622.
URI: http://hdl.handle.net/10119/15877
Material Type: author
Appears in Collections:c10-1. 雑誌掲載論文 (Journal Articles)

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