Adsorption chromatography of proteins

Surovtsev, V. I.; Borzenkov, V. M.; Detushev, K. V.

doi:10.1134/s0006297909020060

Cited by 2 publications

(2 citation statements)

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“…To this end, various chromatographic methods have been developed [ 10 ]. Indeed, many publications reported the description of ion-exchange [ 2 ], size-exclusion [ 8 , 11 ], affinity [ 12 ], and adsorption-based chromatography [ 13 ], as well as their performance and optimal design. Several reviews have also described the fundamental mechanisms, operations, and applications of these chromatographic techniques to that end [ 8 , 14 ].…”

Section: Introductionmentioning

confidence: 99%

Proteomics Methodologies: The Search of Protein Biomarkers Using Microfluidic Systems Coupled to Mass Spectrometry

et al. 2023

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Protein biomarkers have been the subject of intensive studies as a target for disease diagnostics and monitoring. Indeed, biomarkers have been extensively used for personalized medicine. In biological samples, these biomarkers are most often present in low concentrations masked by a biologically complex proteome (e.g., blood) making their detection difficult. This complexity is further increased by the needs to detect proteoforms and proteome complexity such as the dynamic range of compound concentrations. The development of techniques that simultaneously pre-concentrate and identify low-abundance biomarkers in these proteomes constitutes an avant-garde approach to the early detection of pathologies. Chromatographic-based methods are widely used for protein separation, but these methods are not adapted for biomarker discovery, as they require complex sample handling due to the low biomarker concentration. Therefore, microfluidics devices have emerged as a technology to overcome these shortcomings. In terms of detection, mass spectrometry (MS) is the standard analytical tool given its high sensitivity and specificity. However, for MS, the biomarker must be introduced as pure as possible in order to avoid chemical noise and improve sensitivity. As a result, microfluidics coupled with MS has become increasingly popular in the field of biomarker discovery. This review will show the different approaches to protein enrichment using miniaturized devices and the importance of their coupling with MS.

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Section: Introductionmentioning

confidence: 99%

Proteomics Methodologies: The Search of Protein Biomarkers Using Microfluidic Systems Coupled to Mass Spectrometry

et al. 2023

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“…It is well known that the net charge of polypeptides is dependent on the pH, and the acidic or alkaline treatment of a polypeptide can change its net charge. This could result in the electrostatic repulsion between protein layers, consequently leading to the desorption of protein layers. , Therefore, we chose the pH treatment for our investigation of the desorption behavior of enzymatic layers in LbL self-assembled interfaces. This could serve as a basis for investigating an alternative approach to the regeneration of biofunctional interfaces for multiple uses.…”

Section: Introductionmentioning

confidence: 99%

Layer-by-Layer Assembled Carbon Nanotube-Acetylcholinesterase/Biopolymer Renewable Interfaces: SPR and Electrochemical Characterization

et al. 2015

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Developing simple, reliable, and cost-effective methods of renewing an inhibited biocatalyst (e.g., enzymatic interfaces) on biosensors is needed to advance multiuse, reusable sensor applications. We report a method for the renewal of layer-by-layer (LbL) self-assembled inhibition-based enzymatic interfaces in multiwalled carbon nanotube (MWCNT) armored acetylcholinesterase (AChE) biosensors. The self-assembly process of MWCNT dispersed enzymes/biopolymers was investigated using surface plasmon resonance (SPR). The LbL fabrication consisted of alternating cushion layers of positively charged CNT-polyethylenimine (CNT-PEI) and negatively charged CNT-deoxyribonucleic acid (CNT-DNA) and a functional interface consisting of alternating layers of CNT-PEI and negatively charged CNT-acetylcholine esterase (CNT-AChE, pH 7.4). The observed SPR response signal increased while assembling the different layers, indicating the buildup of multiple layers on the Au surface. A partial desorption of the top enzymatic layer in the LbL structure was observed with a desorption strategy employing alkaline treatment. This indicates that the strong interaction of CNT-biopolymer conjugates with the Au surface was a result of both electrostatic interactions between biopolymers and the surface binding energy from CNTs: the closer the layers are to the Au surface, the stronger the interactions. In contrast, a similar LbL assembly of soluble enzyme/polyelectrolytes resulted in stronger desorption on the surface after the alkaline treatment; this led to the investigation of AChE layer removal, permanently inhibited after pesticide exposure on glassy carbon (GC) electrodes, while keeping the cushion layers intact. The desorption strategy permitted the SPR and electrochemical electrode surfaces to be regenerated multiple times by the subsequent self-assembly of fresh PEI/AChE layers. Flow-mode electrochemical amperometric analysis demonstrated good stability toward the determination of acetylcholine with 97.1 ± 2.7% renewability. Our simple, inexpensive approach shows the potential of renewable LbL self-assembled functional interfaces for multiple uses in a wide field of applications such as biosensing, various biotechnological processes, and the food and health industries.

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Adsorption chromatography of proteins

Cited by 2 publications

References 1 publication

Proteomics Methodologies: The Search of Protein Biomarkers Using Microfluidic Systems Coupled to Mass Spectrometry

Proteomics Methodologies: The Search of Protein Biomarkers Using Microfluidic Systems Coupled to Mass Spectrometry

Layer-by-Layer Assembled Carbon Nanotube-Acetylcholinesterase/Biopolymer Renewable Interfaces: SPR and Electrochemical Characterization

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