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Journal of Biomaterials Science, Polymer Edition Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tbsp20
Application of the copolymers containing sulfobetaine methacrylate in protein separation by capillary electrophoresis a
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Fuhu Cao , Lin Tan , Lina Xiang , Songtao Liu & Yanmei Wang
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CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei, 230026, P.R. China. Published online: 02 Aug 2013.
To cite this article: Fuhu Cao, Lin Tan, Lina Xiang, Songtao Liu & Yanmei Wang (2013) Application of the copolymers containing sulfobetaine methacrylate in protein separation by capillary electrophoresis, Journal of Biomaterials Science, Polymer Edition, 24:18, 2058-2070, DOI: 10.1080/09205063.2013.823072 To link to this article: http://dx.doi.org/10.1080/09205063.2013.823072
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &
Downloaded by [University of West Florida] at 13:28 02 January 2015
Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Journal of Biomaterials Science, Polymer Edition, 2013 Vol. 24, No. 18, 2058–2070, http://dx.doi.org/10.1080/09205063.2013.823072
Application of the copolymers containing sulfobetaine methacrylate in protein separation by capillary electrophoresis Fuhu Cao, Lin Tan, Lina Xiang, Songtao Liu and Yanmei Wang*
Downloaded by [University of West Florida] at 13:28 02 January 2015
CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, P.R. China (Received 27 April 2013; accepted 5 July 2013) This study describes the formation of highly efficient antiprotein adsorption random copolymer coating of poly(N,N-dimethylacrylamide-co-sulfobetaine methacrylate) (poly(DMA-co-SBMA)) on the fused-silica capillary inner wall. Firstly, the poly (DMA-co-SBMA)s with different feed ratio (SBMA/DMA) were synthesized via the reversible addition fragmentation chain transfer polymerization. And then, X-ray photoelectron spectroscopy (XPS) and water contact angle (CA) were used to investigate the composition and hydrophilicity of poly(DMA-co-SBMA) coating formed on the glass slide surfaces. CA measurements revealed that the poly(DMAco-SBMA) coating became more hydrophilic with the increment of feed ratio (SBMA/DMA), and at the same time, the XPS results showed that the coating ability was also increased with the increment of feed ratio. Followed, the copolymer was applied to coat the fused-silica capillary inner wall, and the coated capillary was used to separate the mixture of proteins (lysozyme, cytochrome c, ribonuclease A, and α-chymotrypsinogen A) in a pH range from 3.0 to 5.0. Under the optimum conditions, an excellent separation of basic proteins with peak efficiencies ranging from 551,000 to 1509,000 N/m had been accomplished within 10 min. Furthermore, the effect of coating composition on protein separation was also investigated through the comparison of separation efficiency achieved by using bare, PSBMA- and poly (DMA-co-SBMA)-coated capillary, respectively. Keywords: SBMA; coating; protein separation; CE; copolymer
1. Introduction Capillary electrophoresis (CE) has been proven to be an effective method for protein analysis due to its high separation efficiency, high resolution, short analysis time, low sample consumption, and ease of automation.[1–5] Fused-silica is a kind of material that is used to manufacture the capillary owing to its good thermal conductivity and ultraviolet transparency.[6] However, in practice, the efficiencies typically achieved in protein separation are considerably lower due to the protein adsorption produced by the positively charged protein residues and negatively charged silanol groups of the bare fused-silica capillary inner wall. In order to minimize the protein adsorption, numerous approaches have been explored, including the use of extreme pH,[7] zwitterionic [8] and high ionic strength additives.[9] Nevertheless, the most common approach is based on a coating of the capillary inner wall by covalently linked or physically adsorbed *Corresponding author. Email: [email protected] Ó 2013 Taylor & Francis
Downloaded by [University of West Florida] at 13:28 02 January 2015
Journal of Biomaterials Science, Polymer Edition
2059
polymers. Polymers that are physically adsorbed to the capillary inner wall to form a coating have several distinct advantages over covalently linked polymer coatings. These advantages include the simplicity of coating formation, the possibility of coating regeneration, and access to a priori knowledge of coating polymer properties. Several homopolymers have been explored as physically adsorbed coatings.[10–17] Among them, poly(N,N-dimethylacrylamide) (PDMA) exhibits an outstanding self-coating ability on the fused-silica capillary inner wall. But on the other hand, PDMA was ineffective to reduce the protein adsorption due to its hydrophobic interaction with proteins. [13,14] In order to solve the drawback of the protein adsorption on the PDMA coating, a variety of copolymer based on PDMA, such as, PDMA-b-PEO-b-PDMA,[18] hydroxyethylcellulose-graft-PDMA,[19] copolymer consisting of N,N-dimethylacrylamide and allylglycidyl ether (epoxy poly(DMA)),[20] copolymer consisting of N,N-dimethylacrylamide and ethylpyrrolidine methacrylate (DMA-EPyM),[21–23] a brush-like copolymer consisting of poly(ethylene glycol) methyl ether methacrylate and N,N-dimethylacrylamide (PEGMA-DMA) [24] and poly(N,N-dimethylacrylamide-co4-(ethyl)-morpholine methacrylamide) (DMA/MAEM),[25] were used as capillary coating for protein analysis by CE. Although these copolymers can reduce the capillary inner wall–protein interaction, the separation efficiency is unsatisfactory. Therefore, in order to enhance the adsorption ability of the physically adsorbed polymer coatings on the capillary inner wall and the protein seperation efficiency, the development of a kind of polymer which possesses the excellent antibiofouling properties and self-coating ability at the same time still remains active area of research. Jiang and co-workers have reported that poly(sulfobetaine methacrylate) (PSBMA) is highly resistant to non-specific protein adsorption (i.e.
Journal of Biomaterials Science, Polymer Edition Publication details, including instructions for authors and subscription information: http://www.tandfonline.com/loi/tbsp20
Application of the copolymers containing sulfobetaine methacrylate in protein separation by capillary electrophoresis a
a
a
a
Fuhu Cao , Lin Tan , Lina Xiang , Songtao Liu & Yanmei Wang
a
a
CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei, 230026, P.R. China. Published online: 02 Aug 2013.
To cite this article: Fuhu Cao, Lin Tan, Lina Xiang, Songtao Liu & Yanmei Wang (2013) Application of the copolymers containing sulfobetaine methacrylate in protein separation by capillary electrophoresis, Journal of Biomaterials Science, Polymer Edition, 24:18, 2058-2070, DOI: 10.1080/09205063.2013.823072 To link to this article: http://dx.doi.org/10.1080/09205063.2013.823072
PLEASE SCROLL DOWN FOR ARTICLE Taylor & Francis makes every effort to ensure the accuracy of all the information (the “Content”) contained in the publications on our platform. However, Taylor & Francis, our agents, and our licensors make no representations or warranties whatsoever as to the accuracy, completeness, or suitability for any purpose of the Content. Any opinions and views expressed in this publication are the opinions and views of the authors, and are not the views of or endorsed by Taylor & Francis. The accuracy of the Content should not be relied upon and should be independently verified with primary sources of information. Taylor and Francis shall not be liable for any losses, actions, claims, proceedings, demands, costs, expenses, damages, and other liabilities whatsoever or howsoever caused arising directly or indirectly in connection with, in relation to or arising out of the use of the Content. This article may be used for research, teaching, and private study purposes. Any substantial or systematic reproduction, redistribution, reselling, loan, sub-licensing, systematic supply, or distribution in any form to anyone is expressly forbidden. Terms &
Downloaded by [University of West Florida] at 13:28 02 January 2015
Conditions of access and use can be found at http://www.tandfonline.com/page/termsand-conditions
Journal of Biomaterials Science, Polymer Edition, 2013 Vol. 24, No. 18, 2058–2070, http://dx.doi.org/10.1080/09205063.2013.823072
Application of the copolymers containing sulfobetaine methacrylate in protein separation by capillary electrophoresis Fuhu Cao, Lin Tan, Lina Xiang, Songtao Liu and Yanmei Wang*
Downloaded by [University of West Florida] at 13:28 02 January 2015
CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026, P.R. China (Received 27 April 2013; accepted 5 July 2013) This study describes the formation of highly efficient antiprotein adsorption random copolymer coating of poly(N,N-dimethylacrylamide-co-sulfobetaine methacrylate) (poly(DMA-co-SBMA)) on the fused-silica capillary inner wall. Firstly, the poly (DMA-co-SBMA)s with different feed ratio (SBMA/DMA) were synthesized via the reversible addition fragmentation chain transfer polymerization. And then, X-ray photoelectron spectroscopy (XPS) and water contact angle (CA) were used to investigate the composition and hydrophilicity of poly(DMA-co-SBMA) coating formed on the glass slide surfaces. CA measurements revealed that the poly(DMAco-SBMA) coating became more hydrophilic with the increment of feed ratio (SBMA/DMA), and at the same time, the XPS results showed that the coating ability was also increased with the increment of feed ratio. Followed, the copolymer was applied to coat the fused-silica capillary inner wall, and the coated capillary was used to separate the mixture of proteins (lysozyme, cytochrome c, ribonuclease A, and α-chymotrypsinogen A) in a pH range from 3.0 to 5.0. Under the optimum conditions, an excellent separation of basic proteins with peak efficiencies ranging from 551,000 to 1509,000 N/m had been accomplished within 10 min. Furthermore, the effect of coating composition on protein separation was also investigated through the comparison of separation efficiency achieved by using bare, PSBMA- and poly (DMA-co-SBMA)-coated capillary, respectively. Keywords: SBMA; coating; protein separation; CE; copolymer
1. Introduction Capillary electrophoresis (CE) has been proven to be an effective method for protein analysis due to its high separation efficiency, high resolution, short analysis time, low sample consumption, and ease of automation.[1–5] Fused-silica is a kind of material that is used to manufacture the capillary owing to its good thermal conductivity and ultraviolet transparency.[6] However, in practice, the efficiencies typically achieved in protein separation are considerably lower due to the protein adsorption produced by the positively charged protein residues and negatively charged silanol groups of the bare fused-silica capillary inner wall. In order to minimize the protein adsorption, numerous approaches have been explored, including the use of extreme pH,[7] zwitterionic [8] and high ionic strength additives.[9] Nevertheless, the most common approach is based on a coating of the capillary inner wall by covalently linked or physically adsorbed *Corresponding author. Email: [email protected] Ó 2013 Taylor & Francis
Downloaded by [University of West Florida] at 13:28 02 January 2015
Journal of Biomaterials Science, Polymer Edition
2059
polymers. Polymers that are physically adsorbed to the capillary inner wall to form a coating have several distinct advantages over covalently linked polymer coatings. These advantages include the simplicity of coating formation, the possibility of coating regeneration, and access to a priori knowledge of coating polymer properties. Several homopolymers have been explored as physically adsorbed coatings.[10–17] Among them, poly(N,N-dimethylacrylamide) (PDMA) exhibits an outstanding self-coating ability on the fused-silica capillary inner wall. But on the other hand, PDMA was ineffective to reduce the protein adsorption due to its hydrophobic interaction with proteins. [13,14] In order to solve the drawback of the protein adsorption on the PDMA coating, a variety of copolymer based on PDMA, such as, PDMA-b-PEO-b-PDMA,[18] hydroxyethylcellulose-graft-PDMA,[19] copolymer consisting of N,N-dimethylacrylamide and allylglycidyl ether (epoxy poly(DMA)),[20] copolymer consisting of N,N-dimethylacrylamide and ethylpyrrolidine methacrylate (DMA-EPyM),[21–23] a brush-like copolymer consisting of poly(ethylene glycol) methyl ether methacrylate and N,N-dimethylacrylamide (PEGMA-DMA) [24] and poly(N,N-dimethylacrylamide-co4-(ethyl)-morpholine methacrylamide) (DMA/MAEM),[25] were used as capillary coating for protein analysis by CE. Although these copolymers can reduce the capillary inner wall–protein interaction, the separation efficiency is unsatisfactory. Therefore, in order to enhance the adsorption ability of the physically adsorbed polymer coatings on the capillary inner wall and the protein seperation efficiency, the development of a kind of polymer which possesses the excellent antibiofouling properties and self-coating ability at the same time still remains active area of research. Jiang and co-workers have reported that poly(sulfobetaine methacrylate) (PSBMA) is highly resistant to non-specific protein adsorption (i.e.
