Dispersion Mechanism in Gel Chromatography of Proteins

Yamamoto, Shuichi; Nakanishi, Koji; Matsuno, Ryuichi; Kamikubo, Tadashi

doi:10.1080/00021369.1978.10863094

Cited by 3 publications

(3 citation statements)

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“…The exponential gradient behaves similarly to the linear strates the expected behavior, Figure 4. Table IV provides a comparison of the predictions of the present model with experimental result^.^ The theoretical and experimental first moments are in satisfactory agreement, as found previo~sly.~ The second moment comparisons are presented as band widths, which were determined from Figure 3 of Yamamoto et al 4 The second moment (band-broadening) predictions are less accurate when compared to those measured experimentally. As the same sample volume was injected for the three runs of Table IV, the peak width decreases with increasing flow rate.…”

Section: Resultssupporting

confidence: 66%

“…4 have provided experimental data for the type of gradient elution processes discussed in this paper, and so our calculations are based on similar operating conditions. 4 have provided experimental data for the type of gradient elution processes discussed in this paper, and so our calculations are based on similar operating conditions.…”

Section: Resultsmentioning

confidence: 99%

“…The moments for chromatography in a long column yield differential expressions for the normalized moments6 dp, = f(z, tp)dz (3) d~2 = g(z, tp)dz (4) d p 3 = h(z, tp) dz (5) where tp = pi is the temporal position of the peak at the column distance z. These differential expressions for the moments, eqs.…”

Section: Model For Gradient Elutionmentioning

confidence: 99%

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Protein separation by ion exchange chromatography: A model for gradient elution

Kang

McCoy

1989

Biotech & Bioengineering

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Many chromatographic separations involve two stages: (1) an attachment or adsorption step in which solutes to be recovered in purified form are attached to the column matrix, and (2) a change in a property of the solvent, with time, to detach and elute a solute as a separated band. The change of eluant property with time is frequently either a linear gradient or a sequence of (discontinuous) step changes. ' Adsorption, ion exchange, affinity, and hydrophobic chromatography are usually effected by this procedure. For example, in the second stage the solvent composition, ionic strength, or pH may be changed as a function of time. If the solvent property varies linearly with time at the column entrance, then, at constant flow rate, a linear gradient progresses through the column.The output response of the column will depend not only on the type of gradient, but also on the adsorption isotherm. For affinity adsorption the isotherm is strongly irreversible,2 but for ion exchange the isotherm is linear for a range of concentration^.^-^ In the present work, we consider the case of a linear isotherm.Gradient elution is an example of a class of programming and elution techniques used in chromatographic processes. We have applied differential moment theories to study temperature programming,6 velocity programming,' and stationary spatial temperature gradient processes.* These studies considered variation of a property of the column system with either time or column position. Our objective here is to apply this method to more general gradient elutions, where column properties change simultaneously with time and position in the column.Yamamoto and co-worker~~,~ have investigated, theoretically and experimentally, gradient elution in ion exchange chromatography of proteins. By applying a continuousflow plate theory, they considered the effect of linear and stepwise increases of ionic strength on elution characteristics (peak position and width). While their results for the retention time (first moment) are identical to those of the present work, the approach to band-broadening , based on a plate theory, required some simplifying assumptions. co-workers convincingly demonstrate how a simple analysis of small scale experiments can guide the design and operation of large-scale separations of proteins by ion exchange chromatography.33439 There is obvious value in extending and improving these ideas, for example a fundamental treatment of transport phenomena in the column, and by generalizing the technique to other chromatographic separations.In this communication, we 1) extend for the first time the moment theory to gradient elution in ion exchange chromatography; 2) consider linear, exponential, and step gradients; and 3) relate retention time (first moment) and band spreading (second moment) to separation efficiency for gradient elution processes. MODEL FOR GRADIENT ELUTIONThe analysis proposed here generalizes earlier reports ,6-8 which involved column properties varying only as a function of time (temperature or flow program...

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Section: Resultssupporting

confidence: 66%

Section: Resultsmentioning

confidence: 99%