Shaking-Induced Aggregation and Flotation in Immunoglobulin Dispersions: Differences between Water and Water–Ethanol Mixtures

Bunkin, N. F.; Shkirin, A. V.; Ninham, Barry W.; Chirikov, S. N.; Чайков, Л. Л.; Penkov, Nikita V.; Козлов, В. А.; Gudkov, Sergey V.

doi:10.1021/acsomega.0c01444

Cited by 55 publications

(61 citation statements)

References 41 publications

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“…1) There seem to be no any differences between the composition of solutions at very low concentrations (with ultrahigh dilution of the starting substance up to 10 24 times) and water prepared using a similar dilution technique (control water) as the starting substance is no longer found in solutions with very low concentrations. However, as shown in the study [16] where the starting solution contains a protein component, the concentration of the starting components in the solution obtained by serial dilution might not correlate with the estimated concentrations due to flotation. Such solutions may contain residual amounts (traces) of baseline proteins that may become denatured due to dilutions with intensive succussion.…”

Section: Discussionmentioning

confidence: 80%

Analysis of Emission Infrared Spectra of Protein Solutions in Low Concentrations

Penkov

Pen’kova

2020

Front. Phys.

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In this work, based on the method of infrared emission spectroscopy, the study of emission spectra of interferon-gamma (IFNγ) solution in a mixture or surrounded by three low-concentration solutions (IFNγ, antibodies to IFNγ, glycine buffer) or water control was performed. First of all, the solutions of low concentrations themselves were studied. It was shown that low-concentration solutions of IFNγ and antibodies to IFNγ had lower emission intensity in three spectral bands near 800, 1,300 and 2000 cm−1 compared to water control. Glycine buffer solution had a radiation level indistinguishable from that of the control. In this work, the effect of adding these low-concentration solutions to IFNγ (1 mg/ml) was compared to the effect of adding water control to IFNγ. All solutions or water were added in 10% (v/v). It was found that adding each of the three test solutions induced an increase in the radiation intensity of the IFNγ solution in the spectral range of 400–1700 cm−1 (compared to the IFNγ solution with control spike). It was also tested whether the radiation of the studied low-concentration solutions surrounding the IFNγ solution (1 mg/ml) affected the IFNγ radiation. The measurement results were compared to the data obtained for IFNγ surrounded by water control. All three solutions were found to exert a distant effect on the IFNγ solution (1 mg/ml), which was manifested in a decrease in the intensity of its radiation near 1,000 and 1,500 cm−1 compared to the control solution of IFNγ. Thus, the emission spectra of low-concentration aqueous solutions were measured for the first time, and differences in the emission spectra of the IFNγ solution depending on low-concentration additives and the environment were shown. The paper interprets the observed differences and discusses possible mechanisms underlying the observed phenomena.

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Section: Discussionmentioning

confidence: 80%

Analysis of Emission Infrared Spectra of Protein Solutions in Low Concentrations

Penkov

Pen’kova

2020

Front. Phys.

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“…The rationale for this work lies in the published results that have characterized the solutions of HD Abs as a self-organizing dispersed system in which nano-scale objects are generated [55]. It was demonstrated that solution of the HD Abs may contain aggregates of original Abs with gas nanobubbles which survived after multiple dilutions due to the flotation effect [44]. These results are in line with the previously shown presence of nanoparticles of the starting material even at a very high dilution level [56].…”

Section: Discussionmentioning

confidence: 99%

“…First, lactose has changed its physicochemical properties, for example, its conformation, as described in [61] in the process of saturation with highly diluted antibodies (specifically, since such a solution differs from a solution for saturation control). Second, it is reasonable to assume that nanoscale objects based on technologically processed (during dilution and vibration exposure) molecules of the original antibodies are present in the highly diluted antibody solution used for lactose saturation [44]. Apparently, lactose can accumulate these objects during saturation and release them during dissolution.…”

Section: Discussionmentioning

confidence: 99%

“…The estimated theoretical Abs concentration in the final solution was no more than 2.5 × 10 −24 mg/mL, or 2.5 × 10 −18 ppb (w/w). However, it should be noted that the physico-chemical studies showed that for the samples manufactured using the technology of high dilutions such estimation might not be correct [44]. The original substance of the antibodies was produced in accordance with the current EU requirements of Good Manufacturing Practice for starting materials [45] by AB Biotechnology (Edinburgh, UK).…”

Section: Tested Samplesmentioning

confidence: 99%

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Development of Terahertz Time-Domain Spectroscopy for Properties Analysis of Highly Diluted Antibodies

Penkov

Фесенко

2020

Applied Sciences

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This work represents the development of a three-band (Debye relaxation, intermolecular bending, and stretching vibrational bands) approach to the terahertz time-domain spectroscopy (THz-TDS) for the analysis of aqueous solutions of lactose saturated with highly diluted (HD) antibodies to interferon-gamma. Since THz-TDS is sensitive to a very slight changes in water properties, we aimed to assess its applicability for the analysis of HD antibodies. To evaluate water features in tested samples we also used Fourier transform infrared spectroscopy, considering the first overtone of intramolecular stretching vibrations of water molecules. We showed a broadening of the energy distribution of intermolecular bonds in the solution of HD substances in comparison to the control solution. This broadening corresponded to the presence of an active component even in HD samples. The results of both methods were similar; however, with the THz-TDS it was possible to analyze the changes in energy distribution of intermolecular (hydrogen) bonds in the solutions. Therefore, our approach to THz-TDS method is applicable for analyzing structural properties of the solutions containing HD antibodies and for the indirect investigation of altered aqueous properties in water solutions when direct measurements of a substance concentration are not available.

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“…Finally, we studied water samples subjected to intensive shaking on a Vortex MSV-3500 vibrating platform at a frequency of 20 Hz for one minute. These studies were motivated by the results of References [36,37], where it was shown that shaking leads to an increase in the volume number density of nanobubbles by about 10 times. However, after shaking and settling for a certain time, the original properties of water should be restored.…”

Section: The Role Of Shakingmentioning

confidence: 98%

Formation of Water-Free Cavity in the Process of Nafion Swelling in a Cell of Limited Volume; Effect of Polymer Fibers Unwinding

et al. 2020

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When Nafion swells in water, colloidal particles are repelled from the polymer surface; this effect is called the formation exclusion zone (EZ), and the EZ size amounts to several hundred microns. However, still no one has investigated the EZ formation in a cell whose dimension is close to the EZ size. It was also shown that, upon swelling in water, Nafion fibers “unwind” into the water bulk. In the case of a cell of limited volume, unwound fibers abut against the cell windows, and water is completely pushed out from the region between the polymer and the cell window, resulting in a cavity appearance. The temporal dynamics of the collapse of this cavity was studied depending on the cell size. It is shown that the cavity formation occurs due to long-range forces between polymer strands. It turned out that this scenario depends on the isotopic composition of the water, ionic additives and water pretreatment. The role of nanobubbles in the formation and collapse of the cavity were analyzed. The results obtained allowed us to conclude that the EZ formation is precisely due to the unwinding of polymer fibers into the liquid bulk.

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Shaking-Induced Aggregation and Flotation in Immunoglobulin Dispersions: Differences between Water and Water–Ethanol Mixtures

Cited by 55 publications

References 41 publications

Analysis of Emission Infrared Spectra of Protein Solutions in Low Concentrations

Analysis of Emission Infrared Spectra of Protein Solutions in Low Concentrations

Development of Terahertz Time-Domain Spectroscopy for Properties Analysis of Highly Diluted Antibodies

Formation of Water-Free Cavity in the Process of Nafion Swelling in a Cell of Limited Volume; Effect of Polymer Fibers Unwinding

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