Photopolymerized monolithic capillary columns for rapid micro high-performance liquid chromatographic separation of proteins

Dean Lee; Frantisek Svec; Jean M.J. Fréchet

doi:10.1016/j.chroma.2004.04.047

Photopolymerized monolithic capillary columns for rapid micro high-performance liquid chromatographic separation of proteins

Dean Lee, Frantisek Svec, Jean M.J. Fréchet^*

^*Corresponding author for this work

Research output: Contribution to journal › Conference article › peer-review

105 Scopus citations

Abstract

The preparation of monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns using photoinitiated in situ polymerization within 200 μm i.d. capillaries and their application for μHPLC separations of proteins have been studied. The low resistance to flow characteristic of monolithic columns, enabled the use of very high flow rates of up to 100 μL/min representing a flow velocity of 87 mm/s. Very good separations of a model protein mixture consisting of ribonuclease A, cytochrome c, myoglobin, and ovalbumin was achieved in less than 40 s using a very simple single step gradient of the mobile phase. Interestingly, no effect of the pore size on the separations of proteins was observed for these monolithic columns within the size range of 0.66-2.2 μm. The monolithic μHPLC columns are found very robust and no changes in the long term separation performance and back pressure were observed.

Original language	English (US)
Pages (from-to)	53-60
Number of pages	8
Journal	Journal of Chromatography A
Volume	1051
Issue number	1-2
DOIs	https://doi.org/10.1016/j.chroma.2004.04.047
State	Published - Oct 8 2004
Externally published	Yes
Event	17th International Symposium on Microscale Separations - Salzburg, Austria Duration: Feb 8 2004 → Feb 12 2004

Keywords

Monolithic columns
Proteins
Stationary phases, LC

ASJC Scopus subject areas

Analytical Chemistry
Biochemistry
Organic Chemistry

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10.1016/j.chroma.2004.04.047

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title = "Photopolymerized monolithic capillary columns for rapid micro high-performance liquid chromatographic separation of proteins",

abstract = "The preparation of monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns using photoinitiated in situ polymerization within 200 μm i.d. capillaries and their application for μHPLC separations of proteins have been studied. The low resistance to flow characteristic of monolithic columns, enabled the use of very high flow rates of up to 100 μL/min representing a flow velocity of 87 mm/s. Very good separations of a model protein mixture consisting of ribonuclease A, cytochrome c, myoglobin, and ovalbumin was achieved in less than 40 s using a very simple single step gradient of the mobile phase. Interestingly, no effect of the pore size on the separations of proteins was observed for these monolithic columns within the size range of 0.66-2.2 μm. The monolithic μHPLC columns are found very robust and no changes in the long term separation performance and back pressure were observed.",

keywords = "Monolithic columns, Proteins, Stationary phases, LC",

author = "Dean Lee and Frantisek Svec and Fr{\'e}chet, {Jean M.J.}",

year = "2004",

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day = "8",

doi = "10.1016/j.chroma.2004.04.047",

language = "English (US)",

volume = "1051",

pages = "53--60",

journal = "Journal of Chromatography A",

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publisher = "Elsevier",

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note = "17th International Symposium on Microscale Separations ; Conference date: 08-02-2004 Through 12-02-2004",

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AU - Lee, Dean

AU - Svec, Frantisek

AU - Fréchet, Jean M.J.

PY - 2004/10/8

Y1 - 2004/10/8

N2 - The preparation of monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns using photoinitiated in situ polymerization within 200 μm i.d. capillaries and their application for μHPLC separations of proteins have been studied. The low resistance to flow characteristic of monolithic columns, enabled the use of very high flow rates of up to 100 μL/min representing a flow velocity of 87 mm/s. Very good separations of a model protein mixture consisting of ribonuclease A, cytochrome c, myoglobin, and ovalbumin was achieved in less than 40 s using a very simple single step gradient of the mobile phase. Interestingly, no effect of the pore size on the separations of proteins was observed for these monolithic columns within the size range of 0.66-2.2 μm. The monolithic μHPLC columns are found very robust and no changes in the long term separation performance and back pressure were observed.

AB - The preparation of monolithic poly(butyl methacrylate-co-ethylene dimethacrylate) capillary columns using photoinitiated in situ polymerization within 200 μm i.d. capillaries and their application for μHPLC separations of proteins have been studied. The low resistance to flow characteristic of monolithic columns, enabled the use of very high flow rates of up to 100 μL/min representing a flow velocity of 87 mm/s. Very good separations of a model protein mixture consisting of ribonuclease A, cytochrome c, myoglobin, and ovalbumin was achieved in less than 40 s using a very simple single step gradient of the mobile phase. Interestingly, no effect of the pore size on the separations of proteins was observed for these monolithic columns within the size range of 0.66-2.2 μm. The monolithic μHPLC columns are found very robust and no changes in the long term separation performance and back pressure were observed.

KW - Monolithic columns

KW - Proteins

KW - Stationary phases, LC

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U2 - 10.1016/j.chroma.2004.04.047

DO - 10.1016/j.chroma.2004.04.047

M3 - Conference article

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SN - 0021-9673

VL - 1051

SP - 53

EP - 60

JO - Journal of Chromatography A

JF - Journal of Chromatography A

IS - 1-2

T2 - 17th International Symposium on Microscale Separations

Y2 - 8 February 2004 through 12 February 2004

ER -

Photopolymerized monolithic capillary columns for rapid micro high-performance liquid chromatographic separation of proteins

Abstract

Keywords

ASJC Scopus subject areas

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