Cationic lipid vesicles as coating precursors in capillary electrochromatography: Separation of basic proteins and neutral steroids

Bonoli, Matteo; Varjo, Sami; Wiedmer, Susanne Κ.; Riekkola, Marja‐Liisa

doi:10.1016/j.chroma.2005.12.045

Cited by 27 publications

(21 citation statements)

References 40 publications

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“…1,2-Dioleyl-3-trimethylammoniumpropane (DOTAP) lipid vesicles were used for the formation of a semipermanent cationic lipid bilayer in a silica capillary. DOTAP coating was stable for the separation of basic proteins (a-chymotrypsinogen A, ribonuclease A, cytochrome c, lysozyme), with acidic buffers (40 mM acetate buffer at pH 4) [119]. However, the majority of investigations of these types of coating were carried out using drug-like analytes.…”

Section: Physically Attached/adsorbedmentioning

confidence: 99%

Capillary electrochromatography of proteins and peptides

Mikšı́k

Sedláková

2007

J of Separation Science

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This review summarizes applications of CEC for the analysis of proteins and peptides. This "hybrid" technique is useful for the analysis of a broad spectrum of proteins and peptides and is a complementary approach to liquid chromatographic and capillary electrophoretic analysis. All modes of CEC are described--granular packed columns, monolithic stationary phases as well as open-tubular CEC. Attention is also paid to pressurized CEC and the chip-based platform.

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Section: Physically Attached/adsorbedmentioning

confidence: 99%

Capillary electrochromatography of proteins and peptides

Mikšı́k

Sedláková

2007

J of Separation Science

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“…It is necessary to study the effect of pH value of the buffer on separation of proteins because the surface charge and conformational properties of the proteins are strongly dependent on the pH and the morphologies of the copolymers can also be easily tuned by a subtle change of the buffer pH [43,44]. Morphologies and charge of the adsorbed copolymer might also affect the separation of basic proteins [31].…”

Section: Effect Of Buffer Ph On Protein Separationmentioning

confidence: 99%

A novel cationic triblock copolymer as noncovalent coating for the separation of proteins by CE

Cao

Zhu

et al. 2011

Electrophoresis

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A novel noncovalent adsorbed coating for CE has been prepared and explored. This coating was based on quaternized poly(2-(dimethylamino)ethyl methacrylate)-block-poly(ethylene oxide)-block-poly(2-(dimethylamino)ethyl methacrylate) (QDED) triblock copolymer which was synthesized by atomic transfer radical polymerization (ATRP) in our laboratory. The polycationic polymer and the negatively charged fused-silica surface attracted each other through electrostatic interactions and hydrogen bonds. It was demonstrated that the coated capillaries provided an electroosmotic flow with reverse direction, and the magnitude of the electroosmotic flow can be modulated by varying the molecular mass of poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA) block and pH value of the buffer. The effects of the molecular mass of PDMAEMA block in QDED triblock copolymer and pH value of the buffer on the separation of basic proteins were investigated in detail. The triblock copolymer coatings showed higher separation efficiency, better migration time repeatability and would apply to wider range of pH than bare fused-silica capillary when used in separating proteins. Proteins from egg white were also separated through this QDED triblock copolymer-coated capillary. These results demonstrated that the QDED triblock copolymer coatings are suitable for analyzing biosamples.

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“…Vacuum was applied at the outlets of the two microwells to withdraw the liquid into the microwells. The solutions of BSA and DOTAP were allowed to adsorb to the PMMA microwell for 60 min and rinsed by 50 µl of PBS twice and dried with nitrogen [27][28][29][30]. Another flowcell was oxygen plasma treated in 5 min and closed with a COP slide before 50 µl of 50 mM DEGDME was 4!…”

Section: Deposition Of Blocking Agents Into Pmma Microfluidic Flow-cellsmentioning

confidence: 99%

Evaluation of Different Nonspecific Binding Blocking Agents Deposited Inside Poly(methyl methacrylate) Microfluidic Flow-Cells

Gubala

Gandhiraman

et al. 2011

Langmuir

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Poly(methyl methacrylate) (PMMA) flow-cells containing microwells were deposited with different nonspecific binding blocking agents namely bovine serum albumin (BSA), cationic lipid (DOTAP) and diethylene glycol dimethyl ether (DEGDME). Water contact angle (WCA) and atomic force microscope (AFM) measurements were carried out to confirm the successful depositions of BSA, DOTAP, DEGDME onto the PMMA surfaces. Fluorescent intensity measurements were performed to evaluate the degree of non-specific adsorption of Cy5-labeled anti-IgG proteins onto plain and oxygen plasma treated (PT) PMMA flow-cells as well as PMMA flow-cells deposited with different above-mentioned blocking agents. We then employed a label-free detection method called total internal reflection ellipsometry (TIRE) to evaluate the stability of the deposited blocking agents inside the PMMA flow-cells. It was found that while DOTAP was the best agent for blocking the non-specific adsorption, it could be removed from the PMMA surfaces of the flow-cells upon rinsing with phosphate buffer saline (PBS) and later deposited back onto the Au-coated glass sensing substrate of the TIRE. The removal of the blocking agents from PMMA surfaces and their deposition onto the sensing substrate were further manifested by measuring the kinetics and the amount of adsorbed anti-a-hCG proteins. Overall, the dry DEGDME coating by plasma enhanced chemical vapor deposition (PECVD) showed very good blocking and excellent stability for subsequent assay inside the microwells. Our results could be useful when one considers what blocking agents should be used for PMMA-based microfluidic immunosensor or biosensor devices by looking at both the blocking efficiency and the stability of the blocking agent.

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Cationic lipid vesicles as coating precursors in capillary electrochromatography: Separation of basic proteins and neutral steroids

Cited by 27 publications

References 40 publications

Capillary electrochromatography of proteins and peptides

Capillary electrochromatography of proteins and peptides

A novel cationic triblock copolymer as noncovalent coating for the separation of proteins by CE

Evaluation of Different Nonspecific Binding Blocking Agents Deposited Inside Poly(methyl methacrylate) Microfluidic Flow-Cells

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