Chronopotentiometric sensing of native, oligomeric, denatured and aggregated serum albumin at charged surfaces

Římanková, Ludmila; Černocká, Hana; Tihlaříková, Eva; Neděla, Vilém; Ostatná, Veronika

doi:10.1016/j.bioelechem.2022.108100

Cited by 3 publications

(2 citation statements)

References 35 publications

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“…The lyophilized samples were dissolved in distilled water and applied to a lacey carbon film on a copper grid. The solution of star copolymers in water (2 μL) was applied on a TEM grid covered with holey carbon film. , Then, the samples were in situ freeze-dried at 20 °C and 10 Pa in the ESEM specimen chamber (operated under environmental mode). Observation was performed at a beam energy of 30 keV, beam current of 5 pA, and working distance of 5.3 mm in high vacuum mode using a dark-field STEM detector.…”

Section: Methodsmentioning

confidence: 99%

Synthesis and Structure Optimization of Star Copolymers as Tunable Macromolecular Carriers for Minimal Immunogen Vaccine Delivery

Mixová,

Tihlaříková,

Zhu

et al. 2024

Bioconjugate Chem.

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Minimal immunogen vaccines are being developed to focus antibody responses against otherwise challenging targets, including human immunodeficiency virus (HIV), but multimerization of the minimal peptide immunogen on a carrier platform is required for activity. Star copolymers comprising multiple hydrophilic polymer chains (“arms”) radiating from a central dendrimer unit (“core”) were recently reported to be an effective platform for arraying minimal immunogens for inducing antibody responses in mice and primates. However, the impact of different parameters of the star copolymer (e.g., minimal immunogen density and hydrodynamic size) on antibody responses and the optimal synthetic route for controlling those parameters remains to be fully explored. We synthesized a library of star copolymers composed of poly[N-(2-hydroxypropyl)methacrylamide] hydrophilic arms extending from poly(amidoamine) dendrimer cores with the aim of identifying the optimal composition for use as minimal immunogen vaccines. Our results show that the length of the polymer arms has a crucial impact on the star copolymer hydrodynamic size and is precisely tunable over a range of 20–50 nm diameter, while the dendrimer generation affects the maximum number of arms (and therefore minimal immunogens) that can be attached to the surface of the dendrimer. In addition, high-resolution images of selected star copolymer taken by a custom-modified environmental scanning electron microscope enabled the acquisition of high-resolution images, providing new insights into the star copolymer structure. Finally, in vivo studies assessing a star copolymer vaccine comprising an HIV minimal immunogen showed the criticality of polymer arm length in promoting antibody responses and highlighting the importance of composition tunability to yield the desired biological effect.

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Section: Methodsmentioning

confidence: 99%

Synthesis and Structure Optimization of Star Copolymers as Tunable Macromolecular Carriers for Minimal Immunogen Vaccine Delivery

Mixová,

Tihlaříková,

Zhu

et al. 2024

Bioconjugate Chem.

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“…The denaturation of surface-attached proteins can be minimized by adjusting the duration of the protein exposure to the electric field to milliseconds [ 39 ], as well as other experimental conditions, such as solution temperature [ 37 ] and ionic strength [ 40 ]. The high sensitivity of the CPS peak H to structural changes in proteins can be utilized not only for monitoring protein denaturation [ 41 – 43 ], oligomerization and aggregation [ 44 , 45 ], posttranslational modifications [ 46 – 48 ], oxidative damage [ 49 , 50 ], and single- aa replacements [ 51 , 52 ] but also for investigating protein interactions with DNA [ 53 – 55 ], peptides [ 56 ], and other proteins [ 57 – 60 ]. CPS peak H appeared to be particularly useful for analyzing water-soluble and membrane proteins [ 11 , 61 – 65 ].…”

Section: Intrinsic Electroactivity Of Proteinsmentioning

confidence: 99%

Electrochemistry in sensing of molecular interactions of proteins and their behavior in an electric field

Vacek,

Zatloukalová,

Dorčák

et al. 2023

Microchim Acta

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Electrochemical methods can be used not only for the sensitive analysis of proteins but also for deeper research into their structure, transport functions (transfer of electrons and protons), and sensing their interactions with soft and solid surfaces. Last but not least, electrochemical tools are useful for investigating the effect of an electric field on protein structure, the direct application of electrochemical methods for controlling protein function, or the micromanipulation of supramolecular protein structures. There are many experimental arrangements (modalities), from the classic configuration that works with an electrochemical cell to miniaturized electrochemical sensors and microchip platforms. The support of computational chemistry methods which appropriately complement the interpretation framework of experimental results is also important. This text describes recent directions in electrochemical methods for the determination of proteins and briefly summarizes available methodologies for the selective labeling of proteins using redox-active probes. Attention is also paid to the theoretical aspects of electron transport and the effect of an external electric field on the structure of selected proteins. Instead of providing a comprehensive overview, we aim to highlight areas of interest that have not been summarized recently, but, at the same time, represent current trends in the field. Graphical abstract

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Electrochemical analysis of proteins important in cancer: Behavior of anterior gradient receptors, p53 protein and their complexes at charged surfaces

Ostatná

Černocká

Galicová

et al. 2023

Current Opinion in Electrochemistry

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Chronopotentiometric sensing of native, oligomeric, denatured and aggregated serum albumin at charged surfaces

Cited by 3 publications

References 35 publications

Synthesis and Structure Optimization of Star Copolymers as Tunable Macromolecular Carriers for Minimal Immunogen Vaccine Delivery

Synthesis and Structure Optimization of Star Copolymers as Tunable Macromolecular Carriers for Minimal Immunogen Vaccine Delivery

Electrochemistry in sensing of molecular interactions of proteins and their behavior in an electric field

Electrochemical analysis of proteins important in cancer: Behavior of anterior gradient receptors, p53 protein and their complexes at charged surfaces

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