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Uniform magnetic core/shell microspheres functionalized with Ni2+-iminodiacetic acid for one step purification and immobilization of his-tagged enzymes.
ACS Appl Mater Interfaces 2013; 5(7):2626-33AA

Abstract

A facile approach has been developed to synthesize Fe3O4/PMG (poly (N,N'-methylenebisacrylamide-co-glycidyl methacrylate)) core/shell microspheres using distillation-precipitation polymerization. Treating PMG shell with iminodiacetic acid (IDA) and Ni2+ yields composite microspheres of Fe3O4/PMG/IDA-Ni2+. The Ni2+ ions loaded on the surface of microspheres provide abundant docking sites for immobilization of histidine-tagged proteins. The high saturation magnetization of Fe3O4/PMG (23 emu/g), determined by vibrating sample magnetometer (VSM), allows an easy separation of the microspheres from solution under an external magnetic field. The composite microspheres were used to purify two His-tagged cellulolytic enzymes (Cel48F and Cel9G) directly from crude cell lysates with high binding affinity, capacity, and specificity. The microspheres can be recycled for many times without significant loss of binding capacity to enzymes. The immobilized enzymes on the surface of microspheres well retain their biological activities in degradation of cellulose. These materials show great potential in the biomedical and biotechnological applications that require low-cost purification of recombinant proteins and instant enzyme immobilization at an industrial scale.

Authors+Show Affiliations

State Key Laboratory of Molecular Engineering of Polymers, Department of Macromolecular Science, and Laboratory of Advanced Materials, Fudan University, Shanghai 200433, People's Republic of China.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

23470159

Citation

Zhang, Yuting, et al. "Uniform Magnetic Core/shell Microspheres Functionalized With Ni2+-iminodiacetic Acid for One Step Purification and Immobilization of His-tagged Enzymes." ACS Applied Materials & Interfaces, vol. 5, no. 7, 2013, pp. 2626-33.
Zhang Y, Yang Y, Ma W, et al. Uniform magnetic core/shell microspheres functionalized with Ni2+-iminodiacetic acid for one step purification and immobilization of his-tagged enzymes. ACS Appl Mater Interfaces. 2013;5(7):2626-33.
Zhang, Y., Yang, Y., Ma, W., Guo, J., Lin, Y., & Wang, C. (2013). Uniform magnetic core/shell microspheres functionalized with Ni2+-iminodiacetic acid for one step purification and immobilization of his-tagged enzymes. ACS Applied Materials & Interfaces, 5(7), pp. 2626-33. doi:10.1021/am4006786.
Zhang Y, et al. Uniform Magnetic Core/shell Microspheres Functionalized With Ni2+-iminodiacetic Acid for One Step Purification and Immobilization of His-tagged Enzymes. ACS Appl Mater Interfaces. 2013 Apr 10;5(7):2626-33. PubMed PMID: 23470159.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Uniform magnetic core/shell microspheres functionalized with Ni2+-iminodiacetic acid for one step purification and immobilization of his-tagged enzymes. AU - Zhang,Yuting, AU - Yang,Yongkun, AU - Ma,Wanfu, AU - Guo,Jia, AU - Lin,Yao, AU - Wang,Changchun, Y1 - 2013/03/20/ PY - 2013/3/9/entrez PY - 2013/3/9/pubmed PY - 2015/10/16/medline SP - 2626 EP - 33 JF - ACS applied materials & interfaces JO - ACS Appl Mater Interfaces VL - 5 IS - 7 N2 - A facile approach has been developed to synthesize Fe3O4/PMG (poly (N,N'-methylenebisacrylamide-co-glycidyl methacrylate)) core/shell microspheres using distillation-precipitation polymerization. Treating PMG shell with iminodiacetic acid (IDA) and Ni2+ yields composite microspheres of Fe3O4/PMG/IDA-Ni2+. The Ni2+ ions loaded on the surface of microspheres provide abundant docking sites for immobilization of histidine-tagged proteins. The high saturation magnetization of Fe3O4/PMG (23 emu/g), determined by vibrating sample magnetometer (VSM), allows an easy separation of the microspheres from solution under an external magnetic field. The composite microspheres were used to purify two His-tagged cellulolytic enzymes (Cel48F and Cel9G) directly from crude cell lysates with high binding affinity, capacity, and specificity. The microspheres can be recycled for many times without significant loss of binding capacity to enzymes. The immobilized enzymes on the surface of microspheres well retain their biological activities in degradation of cellulose. These materials show great potential in the biomedical and biotechnological applications that require low-cost purification of recombinant proteins and instant enzyme immobilization at an industrial scale. SN - 1944-8252 UR - https://www.unboundmedicine.com/medline/citation/23470159/Uniform_magnetic_core/shell_microspheres_functionalized_with_Ni2+_iminodiacetic_acid_for_one_step_purification_and_immobilization_of_his_tagged_enzymes_ L2 - https://dx.doi.org/10.1021/am4006786 DB - PRIME DP - Unbound Medicine ER -