Poly(methylmethacrylate) Microchip Electrophoresis of Proteins Using Linear-poly(acrylamide) Solutions as Separation Matrix

Okada, Hiroki; Kaji, Noritada; Tokeshi, Manabu; Baba, Yoshinobu

doi:10.2116/analsci.24.321

Cited by 8 publications

(3 citation statements)

References 25 publications

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“…Even though the suppression in EOF has been achieved in the literature, the absolute value of EOF mobility and the degree of suppression have not been reported. Protein separation in PDMA gels has been performed in a PMMA microchip, using a variety of methyl cellulose (MC) compounds as EOF suppressor . In these studies, the concentration of MC was varied from 0.01 to 1% (w/v).…”

Section: Resultsmentioning

confidence: 99%

Surfactant‐induced electroosmotic flow in microfluidic capillaries

Azadi¹,

Tripathi

2012

Electrophoresis

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Control of EOF in microfluidic devices is essential in applications such as protein/DNA sizing and high-throughput drug screening. With the growing popularity of poly(methyl methacrylate) (PMMA) as the substrate for polymeric-based microfludics, it is important to understand the effect of surfactants on EOF in these devices. In this article, we present an extensive investigation exploring changes in EOF rate induced by SDS, polyoxyethylene lauryl ether (Brij35) and CTAB in PMMA microfluidic capillaries. In a standard protein buffer (Tris-Glycine), PMMA capillaries exhibited a cathodic EOF with measured mobility of 1.54 ± 0.1 (× 10⁻⁴ cm²/V.s). In the presence of surfactant below a critical concentration, EOF was independent of surfactant concentration. At high concentrations of surfactants, the electroosmotic mobility was found to linearly increase/decrease as the logarithm of concentration before reaching a constant value. With SDS, the EOF increased by 257% (compared to buffer), while it was decreased by 238% with CTAB. In the case of Brij35, the electroosmotic mobility was reduced by 70%. In a binary surfactant system of SDS/CTAB and SDS/Brij35, addition of oppositely charged CTAB reduced the SDS-induced EOF more effectively compared to nonionic Brij35. We propose possible mechanisms that explain the observed changes in EOF and zeta potential values. Use of neutral polymer coatings in combination with SDS resulted in 50% reduction in the electroosmotic mobility with 0.1% hydroxypropyl methyl cellulose (HPMC), while including 2% poly (N,N-dimethylacrylamide) (PDMA) had no effect. These results will potentially contribute to the development of PMMA-based microfluidic devices.

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Section: Resultsmentioning

confidence: 99%

Surfactant‐induced electroosmotic flow in microfluidic capillaries

Azadi¹,

Tripathi

2012

Electrophoresis

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“…An analysis of two closely related bacteriophages, T2 and T4, showed that this technique was sufficiently sensitive and gave high enough resolution between protein peaks to distinguish these two phages. Baba's group investigated various polymer solutions such as linear‐polyacrylamide 135, and dextran 67 as potential low‐viscosity protein sieving matrices. Reproducible, highly efficient size‐based separation of proteins was successfully achieved with a coated microchip reducing EOF and protein adsorption.…”

Section: Separation Modes For Protein Electrophoresis On Microchipsmentioning

confidence: 99%

Recent innovations in protein separation on microchips by electrophoretic methods: An update

Tran¹,

Ayed²,

Pallandre³

et al. 2009

Electrophoresis

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Lab-on-a-chip electrophoresis is becoming increasingly useful for protein analysis, thanks to recent developments in this field. This review is an update of the review we published at the start of 2008 [Peng, Y., Pallandre, A., Tran, N. T., Taverna, M., Electrophoresis 2008, 29, 156-177]. The superiority of polymers for the manufacture of analytical microchips has been confirmed. This trend implies several modifications to the processes previously used with glass/silicon chips and requires a better understanding of the interfacial phenomena of these materials. Significant progress in chip-based techniques for protein analysis has been made in the last 2 years. In addition to advances in traditional electrokinetic modes, counter-flow gradient focusing techniques have emerged as useful methods not only for separation, but also for the online preconcentration of samples. This review, with more than 175 references, presents recent advances and novel strategies for EOF measurement, surface treatment, sample pretreatment, detection and innovations relating to the different modes of separation.

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“…The lifetimes of these columns were limited (usually less than 10 runs) [12], and the run-to-run reproducibility was poor. Currently, replaceable and water-soluble linear or slightly branched polymers, such as linear polyacrylamide [11–13], poly(ethylene glycol) [11], poly(ethylene oxide) [14], dextran [15–17], pullulan [18, 19] and cross-linked polyacrylamide [20–22] are used as sieving matrices for CGE [5, 11, 23–25]. Availability of these polymer matrices has led to improved reproducibility and robustness of this methodology.…”

Section: Introductionmentioning

confidence: 99%

Protein separation by capillary gel electrophoresis: A review

Zhu

Lü

Liu

2012

Analytica Chimica Acta

177

102

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Capillary gel electrophoresis (CGE) has been used for protein separation for more than two decades. Due to the technology advancement, current CGE methods are becoming more and more robust and reliable for protein analysis, and some of the methods have been routinely used for the analysis of protein-based pharmaceuticals and quality controls. In light of this progress, we survey 147 papers related to CGE separations of proteins and present an overview of this technology. We first introduce briefly the early development of CGE. We then review the methodology, in which we specifically describe the matrices, coatings, and detection strategies used in CGE. CGE using microfabricated channels and incorporation of CGE with two-dimensional protein separations are also discussed in this section. We finally present a few representative applications of CGE for separating proteins in real-world samples.

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Poly(methylmethacrylate) Microchip Electrophoresis of Proteins Using Linear-poly(acrylamide) Solutions as Separation Matrix

Cited by 8 publications

References 25 publications

Surfactant‐induced electroosmotic flow in microfluidic capillaries

Surfactant‐induced electroosmotic flow in microfluidic capillaries

Recent innovations in protein separation on microchips by electrophoretic methods: An update

Protein separation by capillary gel electrophoresis: A review

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