caa a22 4500 606233636 CHVBK 20210128101233.0 cr unu---uuuuu 210128e20150601xx s 000 0 eng 10.1007/s11010-015-2362-x doi (NATIONALLICENCE)springer-10.1007/s11010-015-2362-x Effects of employment of distinct strategies to capture antibody on antibody delivery into cultured cells [Elektronische Daten] [Kana Kuwahara, Kazuki Harada, Ryohei Yamagoshi, Takenori Yamamoto, Yasuo Shinohara] The characteristics of antibody delivery into cultured HeLa cells were examined using two delivery systems. Both systems used a cell-penetrating peptide as a tool for intrusion of an antibody into the cells, but either a "protein A derivative” or "hydrophobic motif” was employed to capture the antibody. When we examined the uptake of the Alexa Fluor-labeled antibody by the use of these two systems, both systems were found to effectively deliver the antibody into the cultured cells. However, when we compared the amount of antibody delivered by these systems with the amount of transferrin uptake, the former was 10 times smaller than the latter. The lower efficiency of antibody delivery than transferrin uptake seemed to be attributable to the involvement of the antibody delivery reagent, which failed to catch the antibody molecule. This interpretation was validated by an experiment using a larger amount of antibody, and the amount of antibody delivered by the "protein A derivative” system under this condition was determined to be 13ng proteins/105cells. The antibody delivery achieved by the "protein A derivative” or "hydrophobic motif” showed two differences, i.e., a difference in intracellular distribution of the delivered antibody molecules and a difference in the fluorescence spectrum observed with cellular lysates. Possible reasons for these differences between the two delivery systems are discussed. Springer Science+Business Media New York, 2015 Antibody delivery nationallicence Cell-penetrating peptide nationallicence Protein A nationallicence Hydrophobic motif nationallicence Kuwahara Kana Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut Harada Kazuki Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut Yamagoshi Ryohei Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut Yamamoto Takenori Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut Shinohara Yasuo Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 404/1-2(2015-06-01), 25-30 0300-8177 404:1-2<25 2015 404 11010 https://doi.org/10.1007/s11010-015-2362-x text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s11010-015-2362-x text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Kuwahara Kana Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut NATIONALLICENCE 700 1- Harada Kazuki Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut NATIONALLICENCE 700 1- Yamagoshi Ryohei Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut NATIONALLICENCE 700 1- Yamamoto Takenori Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut NATIONALLICENCE 700 1- Shinohara Yasuo Institute for Genome Research, University of Tokushima, Kuramoto-cho-3, 770-8503, Tokushima, Japan aut NATIONALLICENCE 773 0- Molecular and Cellular Biochemistry Springer US; http://www.springer-ny.com 404/1-2(2015-06-01), 25-30 0300-8177 404:1-2<25 2015 404 11010