Journal of Nanobiotechnology | |
Ni-modified magnetic nanoparticles for affinity purification of His-tagged proteins from the complex matrix of the silkworm fat body | |
Jirayu Boonyakida1  Kenshin Takemura1  Robert Minkner2  Enoch Y. Park3  Hermann Wätzig4  Jian Xu5  | |
[1] Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan;Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan;Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Beethovenstr. 55, 38106, Braunschweig, Germany;Department of Bioscience, Graduate School of Science and Technology, Shizuoka University, 836 Ohya, Suruga-ku, 422-8529, Shizuoka, Japan;Laboratory of Biotechnology, Green Chemistry Research Division, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Japan;Institute of Medicinal and Pharmaceutical Chemistry, TU Braunschweig, Beethovenstr. 55, 38106, Braunschweig, Germany;Laboratory of Biotechnology, Green Chemistry Research Division, Research Institute of Green Science and Technology, Shizuoka University, 836 Ohya, 422-8529, Shizuoka, Japan;Institute of Biology and Information Science, Biomedical Synthetic Biology Research Center, School of Life Sciences, East China Normal University, 200062, Shanghai, People’s Republic of China; | |
关键词: Magnetic nanoparticles; Affinity purification; Recombinant protein; Silkworm; Magnetic separation; | |
DOI : 10.1186/s12951-020-00715-1 | |
来源: Springer | |
【 摘 要 】
Purification of recombinant proteins is often a challenging matter because high purity and high recovery are desired. If the expressed recombinant protein is also in a complex matrix, such as from the silkworm expression system, purification becomes more challenging. Even if purification from the silkworm expression system is troublesome, it benefits from a high capacity for the production of recombinant proteins. In this study, magnetic nanoparticles (MNPs) were investigated as a suitable tool for the purification of proteins from the complex matrix of the silkworm fat body. The MNPs were modified with nickel so that they have an affinity for His-tagged proteins, as the MNP purification protocol itself does not need special equipment except for a magnet. Among the three different kinds of investigated MNPs, MNPs with sizes of 100 nm to 200 nm and approximately 20 nm-thick nickel shells were the most suitable for our purpose. With them, the total protein amount was reduced by up to at least approximately 77.7%, with a protein recovery of around 50.8% from the silkworm fat body. The minimum binding capacity was estimated to be 83.3 µg protein/mg MNP. Therefore, these MNPs are a promising tool as a purification pretreatment of complex sample matrices.
【 授权许可】
CC BY
【 预 览 】
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