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Ruixia Gao,a Xuan Kong,a Xin Wang,a Xiwen He,a Langxing Chen*a and Yukui Zhang*ab
Received 30th May 2011, Accepted 7th September 2011 DOI: 10.1039/c1jm12414e
C8-modified magnetic NPs have also been applied to separate and enrich peptides for proteomics.19,20 The composite of Fe3O4 magnetic nanocrystals and MCM-41 type periodic mesoporous silica nanospheres was used to separate cytochrome c (Cyt C) from binary Cyt C and bovine serum albumin (BSA) solution.21 Two kinds of silica coated magnetic NPs were used as
This journal is ª The Royal Society of Chemistry 2011
extracted from biological tissues has proven to be very challenging. Therefore, design of highly selective enrichment strategies during sample pretreatment procedures remains an area of intense research interest.
Downloaded by Nankai University on 28 March 2012 Published on 05 October 2011 on http://pubs.rsc.org | doi:10.1039/C1JM12414E
Journal of Materials Chemistry
Several strategies have been developed for the isolation and enrichment of peptides or proteins based on functionalized Fe3O4 NPs with well-defined nanostructures and surface properties. For example, magnetic NPs as immobilized metal affinity chromatography (IMAC) have been used as alternative methods for selective enrichment of various target species.11–18 The enrichment protocols based on IMAC are specific to the proteins with histidine-tagged proteins or histidine/cysteine exposed on the surface of the proteins. However, it has been difficult to separate proteins not possessing histidines or cysteines, which limits the use of this method.
pH-dependent magnetic nano-adsorbents for the selective separation of proteins.22 Although the methods mentioned above are effective for separating protein, the selectivity for the protein of interest has been poor and has not been general. These drawbacks can potentially be solved by molecularly imprinted polymers (MIPs), which can selectively capture a protein in a complex matrix for protein purification.23,24
Βιβλιοθήκη Baidu
PAPER
Preparation and characterization of uniformly sized molecularly imprinted polymers functionalized with core–shell magnetic nanoparticles for the recognition and enrichment of protein
A general method to prepare thin, molecularly imprinted polymer (MIP) coatings on magnetic Fe3O4 nanoparticles (NPs) with a uniform core–shell structure for the recognition and enrichment of protein was developed. Four proteins (bovine serum albumin (BSA, pI ¼ 4.9), bovine hemoglobin (BHb, pI ¼ 6.9), bovine pancreas ribonuclease A (RNase A, pI ¼ 9.4) and lysozyme (Lyz, pI ¼ 11.2)) with different isoelectric points were chosen as the templates. The magnetic protein-MIPs were synthesized by combining surface imprinting and sol–gel techniques. The morphology, adsorption and recognition properties of the magnetic molecularly imprinted NPs were investigated by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy and through the use of a vibrating sample magnetometer (VSM). In comparison with the use of Lyz, BSA and RNase A as template proteins, BHb-imprinted Fe3O4 showed the best imprinting effect and the highest adsorption capacity among the four proteins. The as-prepared Fe3O4@BHb-MIPs NPs with a mean diameter of 230 nm were coated with an MIP shell that was 10 nm thick, which enabled the Fe3O4@BHb-MIPs to easily reach adsorption equilibrium. A high magnetic saturation value of 25.47 emu gÀ1 for Fe3O4@BHb-MIPs NPs was obtained, which endowed the adsorbent with the convenience of magnetic separation under an external magnetic field. The resultant Fe3O4@BHb-MIPs NPs could not only selectively extract a target protein from mixed proteins but also specifically capture the protein BHb from a real sample of bovine blood. In addition, different batches of magnetic MIPs showed good reproducibility and reusability for at least six repeated cycles.
1. Introduction
The selective isolation and detection of protein targets from complex samples is of great significance in key life science disciplines (e.g., diagnostics, proteomics, and protein purification).1,2 Recently, magnetic field-based separations using magnetic nanomaterials have received considerable attention in the fields of biomedicine and biotechnology, including for rapid biological separation, targeted drug delivery, medical imaging and proteomics research.2–10 In particular, magnetic Fe3O4 nanoparticles (NPs) are the most commonly used because of their good biocompatibility, magnetic susceptibility, low toxicity and easy preparation. The key requirement of magnetic Fe3O4 NPs for the enrichment of peptides and proteins is that functionalized Fe3O4 NPs should be specific for their target. In humans, the detection of low-abundance peptides/proteins
J. Mater. Chem., 2011, 21, 17863–17871 | 17863
Downloaded by Nankai University on 28 March 2012 Published on 05 October 2011 on http://pubs.rsc.org | doi:10.1039/C1JM12414E
View Online / Journal Homepage / Table of Contents for this issue
C Dynamic Article Links <
Cite this: J. Mater. Chem., 2011, 21, 17863 www.rsc.org/materials
aDepartment of Chemistry, Nankai University, Tianjin, 300071, P.R. China. E-mail: lxchen@nankai.edu.cn; Fax: +86 22 23502458 bDalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116011, P. R. China. E-mail: ykzhang@dicp.ac.cn
Received 30th May 2011, Accepted 7th September 2011 DOI: 10.1039/c1jm12414e
C8-modified magnetic NPs have also been applied to separate and enrich peptides for proteomics.19,20 The composite of Fe3O4 magnetic nanocrystals and MCM-41 type periodic mesoporous silica nanospheres was used to separate cytochrome c (Cyt C) from binary Cyt C and bovine serum albumin (BSA) solution.21 Two kinds of silica coated magnetic NPs were used as
This journal is ª The Royal Society of Chemistry 2011
extracted from biological tissues has proven to be very challenging. Therefore, design of highly selective enrichment strategies during sample pretreatment procedures remains an area of intense research interest.
Downloaded by Nankai University on 28 March 2012 Published on 05 October 2011 on http://pubs.rsc.org | doi:10.1039/C1JM12414E
Journal of Materials Chemistry
Several strategies have been developed for the isolation and enrichment of peptides or proteins based on functionalized Fe3O4 NPs with well-defined nanostructures and surface properties. For example, magnetic NPs as immobilized metal affinity chromatography (IMAC) have been used as alternative methods for selective enrichment of various target species.11–18 The enrichment protocols based on IMAC are specific to the proteins with histidine-tagged proteins or histidine/cysteine exposed on the surface of the proteins. However, it has been difficult to separate proteins not possessing histidines or cysteines, which limits the use of this method.
pH-dependent magnetic nano-adsorbents for the selective separation of proteins.22 Although the methods mentioned above are effective for separating protein, the selectivity for the protein of interest has been poor and has not been general. These drawbacks can potentially be solved by molecularly imprinted polymers (MIPs), which can selectively capture a protein in a complex matrix for protein purification.23,24
Βιβλιοθήκη Baidu
PAPER
Preparation and characterization of uniformly sized molecularly imprinted polymers functionalized with core–shell magnetic nanoparticles for the recognition and enrichment of protein
A general method to prepare thin, molecularly imprinted polymer (MIP) coatings on magnetic Fe3O4 nanoparticles (NPs) with a uniform core–shell structure for the recognition and enrichment of protein was developed. Four proteins (bovine serum albumin (BSA, pI ¼ 4.9), bovine hemoglobin (BHb, pI ¼ 6.9), bovine pancreas ribonuclease A (RNase A, pI ¼ 9.4) and lysozyme (Lyz, pI ¼ 11.2)) with different isoelectric points were chosen as the templates. The magnetic protein-MIPs were synthesized by combining surface imprinting and sol–gel techniques. The morphology, adsorption and recognition properties of the magnetic molecularly imprinted NPs were investigated by transmission electron microscopy (TEM), scanning electron microscopy (SEM), X-ray diffraction (XRD), and Fourier transform infrared (FT-IR) spectroscopy and through the use of a vibrating sample magnetometer (VSM). In comparison with the use of Lyz, BSA and RNase A as template proteins, BHb-imprinted Fe3O4 showed the best imprinting effect and the highest adsorption capacity among the four proteins. The as-prepared Fe3O4@BHb-MIPs NPs with a mean diameter of 230 nm were coated with an MIP shell that was 10 nm thick, which enabled the Fe3O4@BHb-MIPs to easily reach adsorption equilibrium. A high magnetic saturation value of 25.47 emu gÀ1 for Fe3O4@BHb-MIPs NPs was obtained, which endowed the adsorbent with the convenience of magnetic separation under an external magnetic field. The resultant Fe3O4@BHb-MIPs NPs could not only selectively extract a target protein from mixed proteins but also specifically capture the protein BHb from a real sample of bovine blood. In addition, different batches of magnetic MIPs showed good reproducibility and reusability for at least six repeated cycles.
1. Introduction
The selective isolation and detection of protein targets from complex samples is of great significance in key life science disciplines (e.g., diagnostics, proteomics, and protein purification).1,2 Recently, magnetic field-based separations using magnetic nanomaterials have received considerable attention in the fields of biomedicine and biotechnology, including for rapid biological separation, targeted drug delivery, medical imaging and proteomics research.2–10 In particular, magnetic Fe3O4 nanoparticles (NPs) are the most commonly used because of their good biocompatibility, magnetic susceptibility, low toxicity and easy preparation. The key requirement of magnetic Fe3O4 NPs for the enrichment of peptides and proteins is that functionalized Fe3O4 NPs should be specific for their target. In humans, the detection of low-abundance peptides/proteins
J. Mater. Chem., 2011, 21, 17863–17871 | 17863
Downloaded by Nankai University on 28 March 2012 Published on 05 October 2011 on http://pubs.rsc.org | doi:10.1039/C1JM12414E
View Online / Journal Homepage / Table of Contents for this issue
C Dynamic Article Links <
Cite this: J. Mater. Chem., 2011, 21, 17863 www.rsc.org/materials
aDepartment of Chemistry, Nankai University, Tianjin, 300071, P.R. China. E-mail: lxchen@nankai.edu.cn; Fax: +86 22 23502458 bDalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian, 116011, P. R. China. E-mail: ykzhang@dicp.ac.cn