Ni-modified magnetic nanoparticles for affinity purification of His-tagged proteins from the complex matrix of the silkworm fat body

Journal of Nanobiotechnology

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