Capillary electrophoresis‐mass spectrometry for intact protein analysis: Pharmaceutical and biomedical applications (2018–March 2023)

Maráková, Katarína; Opetová, Martina; Tomašovský, Radovan

doi:10.1002/jssc.202300244

Cited by 19 publications

(4 citation statements)

References 266 publications

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“…Biopharmaceuticals may contain multiple species of various sizes, such as product-related aggregates (dimers, tetramers), and fragments of low molecular mass relative to the parent molecule. CE-MS enables to determination of the relative molecular masses of these species and thus allows evaluation of the purity of biopharmaceutical products, as well as their close monitoring during the production process and storage [89,90].…”

Section: 62mentioning

confidence: 99%

Determination of physicochemical parameters of (bio)molecules and (bio)particles by capillary electromigration methods

Štěpánová,

Kašička

2024

J of Separation Science

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The review provides an overview of recent developments and applications of capillary electromigration (CE) methods for the determination of important physicochemical parameters of various (bio)molecules and (bio)particles. These parameters include actual and limiting (absolute) ionic mobilities, effective electrophoretic mobilities, effective charges, isoelectric points, electrokinetic potentials, hydrodynamic radii, diffusion coefficients, relative molecular masses, acidity (ionization) constants, binding constants and stoichiometry of (bio)molecular complexes, changes of Gibbs free energy, enthalpy and entropy and rate constants of chemical reactions and interactions, retention factors and partition and distribution coefficients. For the determination of these parameters, the following CE methods are employed: zone electrophoresis in a free solution or in sieving media, isotachophoresis, isoelectric focusing, affinity electrophoresis, electrokinetic chromatography, and electrochromatography. In the individual sections, the procedures for the determination of the above parameters by the particular CE methods are described.

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Section: 62mentioning

confidence: 99%

Determination of physicochemical parameters of (bio)molecules and (bio)particles by capillary electromigration methods

Štěpánová,

Kašička

2024

J of Separation Science

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“…CE, as an electric field-mediated separation technique, was first introduced in 1981. It is now widely used to analyze molecules of different sizes in various applications, outperforming or complementing liquid chromatographic techniques [ 114 , 115 ]. CE is a cost-effective instrument that requires minimal reagents and offers high separation efficiency [ 116 ].…”

Section: Analysis Of Drug Residues In Foodmentioning

confidence: 99%

Recent Advances in the Determination of Veterinary Drug Residues in Food

Pratiwi,

Ramadhanti,

Amatulloh

et al. 2023

Foods

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The presence of drug residues in food products has become a growing concern because of the adverse health risks and regulatory implications. Drug residues in food refer to the presence of pharmaceutical compounds or their metabolites in products such as meat, fish, eggs, poultry and ready-to-eat foods, which are intended for human consumption. These residues can come from the use of drugs in the field of veterinary medicine, such as antibiotics, antiparasitic agents, growth promoters and other veterinary drugs given to livestock and aquaculture with the aim of providing them as prophylaxis, therapy and for promoting growth. Various analytical techniques are used for this purpose to control the maximum residue limit. Compliance with the maximum residue limit is very important for food manufacturers according to the Food and Drug Administration (FDA) or European Union (EU) regulations. Effective monitoring and control of drug residues in food requires continuous advances in analytical techniques. Few studies have been reviewed on sample extraction and preparation techniques as well as challenges and future directions for the determination of veterinary drug residues in food. This current review focuses on the overview of regulations, classifications and types of food, as well as the latest analytical methods that have been used in recent years (2020–2023) for the determination of drug residues in food so that appropriate methods and accurate results can be used. The results show that chromatography is still a widely used technique for the determination of drug residue in food. Other approaches have been developed including immunoassay, biosensors, electrophoresis and molecular-based methods. This review provides a new development method that has been used to control veterinary drug residue limit in food.

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“…Folding/unfolding studies using either chemical denaturant or temperature have shown a significant distinction in the μ between folded, partially unfolded and unfolded populations. [18][19][20][21][22][23] Studies have interfaced CE with intact protein mass spectrometry (CE-MS) [24][25] to unambiguously assign different peaks related to the same protein, confirming the separation of conformers or post-translationally modified isoforms. [26][27] A folding transition has also been monitored using CE and has been validated through comparison with ion-mobility MS (IM-MS) which is sensitive to conformational differences via changes in protein cross-sectional area.…”

Section: Introductionmentioning

confidence: 99%

Using Capillary Electrophoresis to Investigate Protein Conformational and Compositional Heterogeneity

Grosas,

Du Plessis,

Thevarajah

et al. 2024

ChemBioChem

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Detailed insights into protein structure/function relationships require robust characterization methodologies. Free‐solution capillary electrophoresis (CE) is a unique separation technique which is sensitive to the conformation and/or composition of proteins, and therefore provides information on the heterogeneity of these properties. Three unrelated, conformationally/compositionally‐altered proteins were separated by CE. An electrophoretic mobility distribution was determined for each protein along with its conformational and/or compositional heterogeneity. CE results were compared with molar mass distributions obtained from size‐exclusion chromatography coupled to light scattering (SEC‐MALS). Bovine serum albumin oligomers and two monomeric species were separated, highlighting variations in conformational/compositional heterogeneity among the oligomers. Analysis of yeast alcohol dehydrogenase resolved two monomeric conformers and various tetrameric species, illustrating the impact of zinc ion removal and disulfide bond reduction on the protein’s heterogeneity. The apo (calcium‐free) and holo forms of bovine α‐lactalbumin were separated and differences in the species’ heterogeneity was measured; by contrast, the SEC‐MALS profiles were identical. Comparative analysis of these structurally unrelated proteins provided novel insights into the interplay between molar mass and conformational/compositional heterogeneity. Overall, this study expands the utility of CE by demonstrating its capacity to discern protein species and their heterogeneity, properties which are not readily accessible by other analytical techniques.

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Capillary electrophoresis‐mass spectrometry for intact protein analysis: Pharmaceutical and biomedical applications (2018–March 2023)

Cited by 19 publications

References 266 publications

Determination of physicochemical parameters of (bio)molecules and (bio)particles by capillary electromigration methods

Determination of physicochemical parameters of (bio)molecules and (bio)particles by capillary electromigration methods

Recent Advances in the Determination of Veterinary Drug Residues in Food

Using Capillary Electrophoresis to Investigate Protein Conformational and Compositional Heterogeneity

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