Molecular-sieve chromatography and electrophoresis in polyacrylamide gels

Morris, C. J. O. R.; Morris, Peggy

doi:10.1042/bj1240517

Cited by 68 publications

(24 citation statements)

References 19 publications

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“…In the earliest model, the so-called Ogston-model [20,21], the gel is assumed to act as a sieve with a distribution of pore sizes and the separation is regarded as a kind of filtration, driven by the electric field. This leads to a mobility m proportional to exp(±conc.…”

Section: Mechanism Of Dna Movementmentioning

confidence: 99%

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Heller

1999

Electrophoresis

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We have studied the separation of single‐stranded and double‐stranded DNA in a matrix of entangled, linear poly‐N,N‐dimethylacrylamide. Our results give better insight into the mechanisms involved during separations in polymer solutions. The dependence of different parameters on DNA size, electric field, pore size and the polymer chain length are evaluated and compared to theoretical predictions. Striking differences between experimental data and predicted scaling laws are found. Our data should help to optimize DNA separation in capillary electrophoresis and to improve existing models for DNA separation in porous matrices.

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Section: Mechanism Of Dna Movementmentioning

confidence: 99%

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Heller

1999

Electrophoresis

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“…Diffusive transport of large and small solutes in hydrogels has been studied by gel permeation chromatography (27)(28)(29), diffusion cells (30), light scattering (31,32,33), pulsed field gradient NMR (34 -37), and other methods (38). Diffusion of solutes in hydrogels is expected to depend upon the size of the probe relative to the size of the pore spaces in the gel, the geometry of the gel structure, and various physical/chemical interactions between the probe species and the gel matrix.…”

Section: Introductionmentioning

confidence: 99%

“…Diffusion of solutes in hydrogels is expected to depend upon the size of the probe relative to the size of the pore spaces in the gel, the geometry of the gel structure, and various physical/chemical interactions between the probe species and the gel matrix. In order to account for probe size, the Ogston theory has been extended to diffusion (and electrophoresis) (27,28,39) whereby the partition coefficient has been equated to the ratio of the diffusion coefficient in the gel to that in free solution. Other approaches, including hindered transport theory (40,41), have also been used to describe the effects of solute size on diffusion.…”

Section: Introductionmentioning

confidence: 99%

Proton Diffusion andT1Relaxation in Polyacrylamide Gels: A Unified Approach Using Volume Averaging

Penke

Kinsey

Gibbs

et al. 1998

Journal of Magnetic Resonance

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The structure of polyacrylamide gels was studied using proton spin-lattice relaxation and PFG diffusion methods. Polyacrylamide gels, with total polymer concentrations ranging from 0.25 to 0.35 g/ml and crosslinker concentrations from 0 to 10% by weight, were studied. The data showed no effect of the crosslinker concentration on the diffusion of water molecules. The Ogston-Morris and Mackie-Meares models fit the general trends observed for water diffusion in gels. The diffusion coefficients from the volume averaging method also fit the data, and this theory was able to account for the effects of water-gel interactions that are not accounted for in the other two theories. The averaging theory also did not require the physically unrealistic assumption, required in the other two theories, that the acrylamide fibers are of similar size to water molecules. Contrary to the diffusion data, T 1 relaxation measurements showed a significant effect of crosslinker concentration on the relaxation of water in gels. The model developed using the Bloch equations and the volume averaging method described the effects of water adsorption on the gel medium on both the diffusion coefficients and the relaxation measurements. In the proposed model the gel medium was assumed to consist of three phases (i.e., bulk water, uncrosslinked acrylamide fibers, and a bisacrylamide crosslinker phase). The effects of the crosslinker concentration were accounted for by introducing the proton partition coefficient, K eq , between the bulk water and crosslinker phase. The derived relaxation equations were successful in fitting the experimental data. The partition coefficient, K eq , decreased significantly as the crosslinker concentration increased from 5 to 10% by weight. This trend is consistent with the idea that bisacrylamide tends to form hydrophobic regions with increasing crosslinker concentration.

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“…The apparent size and the shape of the polymers in solution is the basis for separation in size exclusion chromatography [8,9]. This simple separation mechanism relies on protein size differences, packing material pore size and particle size as well as the operating conditions such as sample concentration and volume for the successful resolution of protein mixtures [10].…”

Section: Introductionmentioning

confidence: 99%

“…Size exclusion packing material has evolved from the large particle polysaccharide soft gels to small, rigid packing that offers enhanced resolution and significantly faster separation [11][12][13]. The principle feature of SEC is its gentle non-adsorptive interaction with the sample, enabling high retention of biomolecular activity [8,10].…”

Section: Introductionmentioning

confidence: 99%

Gel permeation chromatographic method for drug protein binding studies

Bhatia¹,

Jadhav²,

Kumbhar³

2017

mpj

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The estimation of individual plasma protein in free and bound form with analytes has importance in pharmacokinetics study. Albumins and globulins are most abundantly found in plasma and plays crucial role in plasma protein binding. The present communication deals with the development of gel permeation chromatographic method for the estimation of plasma protein binding studies for ketoprofen, tapentadol and furaltadone. The method was developed using WATER system with Ultrahydrogel column (7.8 x 300 mm i.d.), refractive index detector (1.00 to 1.75 RIU) and Rheodyne injection valve fitted with a 20µl sample loop using 0.1% sodium nitrate as a mobile phase. The method was developed by studying binding above drugs with most of plasma proteins. These complexes have shown linearity in range of 100 to 300µg/ml of these drugs. The developed method has been validated using USFDA guidelines. The developed bioanalytical method is accurate, precise, and selective and sensitive for quantification of plasma proteins bound drugs. The further optimization of method using other standard plasma proteins will explore its applicability in pharmacokinetic and biopharmaceutical studies of most of drugs.

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Molecular-sieve chromatography and electrophoresis in polyacrylamide gels

Cited by 68 publications

References 19 publications

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Separation of double-stranded and single-stranded DNA in polymer solutions: I. Mobility and separation mechanism

Proton Diffusion andT1Relaxation in Polyacrylamide Gels: A Unified Approach Using Volume Averaging

Gel permeation chromatographic method for drug protein binding studies

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