Vizualization of protein retention and migration in reversed-phase liquid chromatography

Bussolo, Joseph M. Di; Gant, J.R.

doi:10.1016/s0021-9673(01)81638-1

Cited by 20 publications

(4 citation statements)

References 8 publications

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“…While protein migration has been extensively investigated in phase liquid chromatography [ 16 ] or under conditions where an external force is applied (e.g., shear forces under flow), [ 17 ] the effect of passive migration on the functionality of immobilized proteins has not yet been explored. Protein migration triggered by either storage or operational conditions may alter the protein distribution along the support surface and indirectly affect the functional properties of the immobilized proteins.…”

Section: Introductionmentioning

confidence: 99%

Intraparticle Macromolecular Migration Alters the Structure and Function of Proteins Reversibly Immobilized on Porous Microbeads

Diamanti

Arana-Peña

Comino

et al. 2022

Adv Materials Inter

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While migration of reversibly immobilized proteins across the volume of supports is investigated in conditions where an external force is applied or under fluid flow conditions, their passive migration upon sample storage and its effect on the protein functionality remain unexplored. Understanding such intraparticle macromolecular migration is essential to develop protein functionalized biomaterials with a longer life span. This work investigates the spatiotemporal migration of His‐tagged immobilized fluorescent proteins inside porous agarose microbeads under different storage conditions. A tool that assesses the intraparticle protein migration across the surface of the porous supports is developed. Differences in migration patterns between different proteins suggest that binding dynamics between proteins and their supports play a key role in their migration. The effect of macromolecular migration on the functional and structural properties of bound proteins and enzymes is also explored. Therefore, single‐particle measurements to understand how the migration process affects the functionality of immobilized enzymes are performed. Evaluating protein migration and understanding the reason behind such phenomena allows gaining control over immobilization processes and design immobilization chemistries that either prevent or promote intraparticle macromolecular diffusion upon storage, depending on the desired final application.

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Section: Introductionmentioning

confidence: 99%

Intraparticle Macromolecular Migration Alters the Structure and Function of Proteins Reversibly Immobilized on Porous Microbeads

Diamanti

Arana-Peña

Comino

et al. 2022

Adv Materials Inter

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“…11 In an experimental ap-proach, Bussolo and Gant discussed the visualization of protein retention and migration in RP liquid chromatography. 12 Ilg et al tried to observe the band profiles by magnetic resonance imaging and discussed the wall effect of a preparative column. 13 To get a clearer visualization of the problem, we developed a video image analyzing system for liquid chromatography.…”

mentioning

confidence: 99%

“…Many theoretical approaches have been developed to clarify the band migration and the separation process. − Gelderloos et al showed the necessity and usefulness of whole-column detection with computer simulations . In an experimental approach, Bussolo and Gant discussed the visualization of protein retention and migration in RP liquid chromatography . Ilg et al tried to observe the band profiles by magnetic resonance imaging and discussed the wall effect of a preparative column …”

mentioning

confidence: 99%

Direct Imaging of the Stepwise Elution Process in High-Performance Liquid Chromatography

Tamura

Razee

et al. 1996

Anal. Chem.

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“…In addition, the use of glass analytical columns allows the separation to be visually monitored, simplifying optimization of the plug composition and providing immediate information on the number and nature of sample components. Previously employed to follow the retention of various proteins in reverse-phase LC (26), the transparent glass columns used in the reported work permitted the equilibration of the various dyes between the stationary and the mobile phases to be followed in some detail, yielding a great deal of chemical information. In addition, there is the potential for direct on-column detection (i.e., using laser scanning) along the entire column.…”

mentioning

confidence: 99%

Transitory mobile phase environments for rapid selectivity changes in liquid chromatography: application to organic dyestuffs

Gluckman¹,

Šlais²,

Brinkman³

et al. 1987

Anal. Chem.

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Transient mobile phase environments created by in|ectlng solvent plugs of varying composition are shown to selectively elute compounds according to their chemical properties. With several organic dyestuffs as model solutes, weakly acidic, strongly acidic, and neutral substances were sequentially eluted In a series of subchromatograms based on pH, Ion-pair competition, and hydrophobic Interaction mechanisms, respectively. Elution with both Isocratlc plugs and with pluggenerated gradients Is described. The use of transparent columns allowed solute bands to be viewed during the separation process, and the potential for on-column detection and laser scanning Is discussed.

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Vizualization of protein retention and migration in reversed-phase liquid chromatography

Cited by 20 publications

References 8 publications

Intraparticle Macromolecular Migration Alters the Structure and Function of Proteins Reversibly Immobilized on Porous Microbeads

Intraparticle Macromolecular Migration Alters the Structure and Function of Proteins Reversibly Immobilized on Porous Microbeads

Direct Imaging of the Stepwise Elution Process in High-Performance Liquid Chromatography

Transitory mobile phase environments for rapid selectivity changes in liquid chromatography: application to organic dyestuffs

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