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Synthesis of micron-sized magnetic agarose beads chelated with nickel ions towards the affinity-based separation of histidine-tagged/rich proteins.
Zhao, Ya-Qi; Yu, Shi-Song; Chen, Meng-Ying; Wang, Yuan; Shi, Yu-Jun; Wang, Xin-Yu; Zhao, Jia-Meng; Dong, Lin-Yi; Zhao, Zhen-Yu; Wang, Xian-Hua.
Afiliação
  • Zhao YQ; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital /
  • Yu SS; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital /
  • Chen MY; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital /
  • Wang Y; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China.
  • Shi YJ; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital /
  • Wang XY; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China.
  • Zhao JM; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital /
  • Dong LY; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China. Electronic address: donglinyi@tmu.edu.cn.
  • Zhao ZY; NHC Key Laboratory of Hormones and Development / Tianjin Key Laboratory of Metabolic Diseases, Chu Hsien-I Memorial Hospital / Tianjin Institute of Endocrinology, Tianjin Medical University, Tianjin 300134, China. Electronic address: zhaozhenyu0858@163.com.
  • Wang XH; Tianjin Key Laboratory on Technologies Enabling Development of Clinical Therapeutics and Diagnostics, School of Pharmacy, Tianjin Medical University, Tianjin 300070, China. Electronic address: xianhua.w@163.com.
J Chromatogr A ; 1708: 464365, 2023 Oct 11.
Article em En | MEDLINE | ID: mdl-37696128
ABSTRACT
Developing high-performance magnetic particles for the effective separation and purification of target proteins has become an important topic in the area of biomedical research. In this work, a simple and novel strategy was proposed for fabricating magnetic Fe3O4@agarose-iminodiacetic acid-Ni microspheres (MAIN), which can efficiently and selectively isolate histidine-tagged/rich proteins (His-proteins). Based on the thermoreversible sol-gel transition of agarose, basic magnetic agarose microspheres were prepared through the inverse emulsion method, in which the emulsion contained agarose and amine-modified Fe3O4 nanoparticles. The size of the emulsion was controlled by the emulsification of a high-speed shear machine, which improved the specific surface area of MAIN. Subsequently, the amine-modified Fe3O4 nanoparticles were covalently crosslinked with agarose through epichlorohydrin, which could avoid leakage of the magnetic source during use and increase the stability of MAIN. The microsized MAIN exhibited a clearly visible spherical core-shell structure with a diameter range from 3.4 µm to 9.8 µm, and excellent suspension ability in aqueous solution. The maximum adsorption capacity of MAIN for histidine-rich bovine hemoglobin was 1069.2 mg g-1 at 35 °C, which was higher than those of commercialized and most reported magnetic agarose microspheres/nanoparticles. The MAIN showed excellent adsorption ability and selectivity toward His-proteins in a mixture of histidine-rich bovine serum albumin (BSA) and histidine-poor lysozyme (LYZ). When the amount of LYZ was 5-fold higher than that of BSA, the recovery of BSA reached 75.0%. To prove its practicability, MAIN was successfully employed for the enrichment of histidine-tagged RSV-F0 from the cell culture medium supernatant. According to the optimized conditions, MAIN could enrich approximately 0.1 mg of RSV-F0 from 1 mL of complex biological sample. Therefore, we believe that the novel MAIN could be applicable for efficient separation and purification of His-proteins from complex biological systems.
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Texto completo: 1 Bases de dados: MEDLINE Assunto principal: Histidina / Níquel Idioma: En Revista: J Chromatogr A Ano de publicação: 2023 Tipo de documento: Article

Texto completo: 1 Bases de dados: MEDLINE Assunto principal: Histidina / Níquel Idioma: En Revista: J Chromatogr A Ano de publicação: 2023 Tipo de documento: Article