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Nanochannel pH Gradient Electrofocusing of Proteins.

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Title: Nanochannel pH Gradient Electrofocusing of Proteins.
Authors: Startsev, Michael A.1 michael.startsev@mq.edu.au, Inglis, David W.2, Baker, Mark S.3, Goldys, Ewa M.1
Source: Analytical Chemistry. 8/6/2013, Vol. 85 Issue 15, p7133-7138. 6p.
Subjects: Nanostructures, pH gradients, Isoelectric focusing, Proteins, Nanofluidics, Mass spectrometry, Buffer action (Chemistry), Electro-osmosis
Abstract: We demonstrate matrix-free pH gradient electrofocusing of proteins within an 85 nm deep nanochannel. In contrast to conventional isoelectric focusing where the fluid does not move, this pH gradient method traps protein molecules flowing through a channel by balancing electric forces due to pH-dependent protein charge and viscous drag forces caused by electro-osmosis. The nanoscale depth of the device and the low voltage used limit convection relative to diffusion, thus producing a stable focused band of protein. R-Phycoerythrin (RPE) and Dylight labeled streptavidin (Dyl-Strep) were focused within a nanochannel using applied voltages between 0.4 and 1.6 V. Concentration enhancement factors of over 380 have been achieved within 5 min. Varying the buffer pH (between 2.7 and 7.2) at the boundaries of the nanochannel affected the shape of the focused bands. For RPE, a pH span of 4.5 (pH 2.7 to 7.2) yielded the narrowest peak while a span of 2.4 (pH 2.7 to 5.1) produced a significantly wider peak. Such matrix-free nanofluidic devices with pH gradient electrofocusing may enable on-chip integration of orthogonal separation techniques with mass spectrometry offering labor savings and enhanced performance. [ABSTRACT FROM AUTHOR]
Copyright of Analytical Chemistry is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Nanochannel pH Gradient Electrofocusing of Proteins.
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  Data: <searchLink fieldCode="JN" term="%22Analytical+Chemistry%22">Analytical Chemistry</searchLink>. 8/6/2013, Vol. 85 Issue 15, p7133-7138. 6p.
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  Data: <searchLink fieldCode="DE" term="%22Nanostructures%22">Nanostructures</searchLink><br /><searchLink fieldCode="DE" term="%22pH+gradients%22">pH gradients</searchLink><br /><searchLink fieldCode="DE" term="%22Isoelectric+focusing%22">Isoelectric focusing</searchLink><br /><searchLink fieldCode="DE" term="%22Proteins%22">Proteins</searchLink><br /><searchLink fieldCode="DE" term="%22Nanofluidics%22">Nanofluidics</searchLink><br /><searchLink fieldCode="DE" term="%22Mass+spectrometry%22">Mass spectrometry</searchLink><br /><searchLink fieldCode="DE" term="%22Buffer+action+%28Chemistry%29%22">Buffer action (Chemistry)</searchLink><br /><searchLink fieldCode="DE" term="%22Electro-osmosis%22">Electro-osmosis</searchLink>
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  Data: We demonstrate matrix-free pH gradient electrofocusing of proteins within an 85 nm deep nanochannel. In contrast to conventional isoelectric focusing where the fluid does not move, this pH gradient method traps protein molecules flowing through a channel by balancing electric forces due to pH-dependent protein charge and viscous drag forces caused by electro-osmosis. The nanoscale depth of the device and the low voltage used limit convection relative to diffusion, thus producing a stable focused band of protein. R-Phycoerythrin (RPE) and Dylight labeled streptavidin (Dyl-Strep) were focused within a nanochannel using applied voltages between 0.4 and 1.6 V. Concentration enhancement factors of over 380 have been achieved within 5 min. Varying the buffer pH (between 2.7 and 7.2) at the boundaries of the nanochannel affected the shape of the focused bands. For RPE, a pH span of 4.5 (pH 2.7 to 7.2) yielded the narrowest peak while a span of 2.4 (pH 2.7 to 5.1) produced a significantly wider peak. Such matrix-free nanofluidic devices with pH gradient electrofocusing may enable on-chip integration of orthogonal separation techniques with mass spectrometry offering labor savings and enhanced performance. [ABSTRACT FROM AUTHOR]
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  Data: <i>Copyright of Analytical Chemistry is the property of American Chemical Society and its content may not be copied or emailed to multiple sites without the copyright holder's express written permission. Additionally, content may not be used with any artificial intelligence tools or machine learning technologies. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.</i> (Copyright applies to all Abstracts.)
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        Value: 10.1021/ac4014447
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        Text: English
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        Type: general
      – SubjectFull: pH gradients
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      – SubjectFull: Isoelectric focusing
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      – SubjectFull: Proteins
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      – SubjectFull: Nanofluidics
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      – SubjectFull: Mass spectrometry
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      – SubjectFull: Buffer action (Chemistry)
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      – SubjectFull: Electro-osmosis
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      – TitleFull: Nanochannel pH Gradient Electrofocusing of Proteins.
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              Text: 8/6/2013
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              Y: 2013
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