Antibodies Processed Using High Dilution Technology Distantly Change Structural Properties of IFNγ Aqueous Solution

Penkov, Nikita. "Antibodies Processed Using High Dilution Technology Distantly Change Structural Properties of IFNγ Aqueous Solution." Pharmaceutics 13, no. 11 (2021): 1864.

for full paper see https://www.mdpi.com/1999-4923/13/11/1864

Abstract

Terahertz spectroscopy allows for the analysis of vibrations corresponding to the large-scale structural movements and collective dynamics of hydrogen-bonded water molecules. Previously, differences had been detected in the emission spectra of interferon-gamma (IFNγ) solutions surrounded by extremely diluted solutions of either IFNγ or antibodies to IFNγ without direct contact compared to a control. Here we aimed to analyse the structural properties of water in a sample of an aqueous solution of IFNγ via terahertz time-domain spectroscopy (THz-TDS). Tubes with the IFNγ solution were immersed in fluidised lactose saturated with test samples (dilutions of antibodies to IFNγ or control) and incubated at 37 ◦C for 1, 1.5–2, 2.5–3, or 3.5–4 h. Fluidised lactose was chosen since it is an excipient in the manufacture of drugs based on diluted antibodies to IFNγ. After incubation, spectra were recorded within a wavenumber range of 10 to 110 cm−1 with a resolution of 4 cm−1 . Lactose saturated with dilutions of antibodies to IFNγ (incubated for more than 2.5 h) changed the structural properties of an IFNγ aqueous solution without direct contact compared to the control. Terahertz spectra revealed stronger intermolecular hydrogen bonds and an increase in the relaxation time of free and weakly bound water molecules. The methodology developed on the basis of THz-TDS could potentially be applied to quality control of pharmaceuticals based on extremely diluted antibodies.

Experimental 

2.2. THz-TDS 

In this study, we used the THz-TDS technique. It allows the simultaneous acquisition of absorption and refraction spectra of the test substance to calculate complex dielectric functions without using the Kramers–Kronig transformations. The details of this method are described, for example, in [46]. The spectra were recorded with a TPS Spectra 3000spectrometer (Teraview, Cambridge, UK) in a wavenumber range from 10 to 110 cm−1 with a resolution of 4 cm−1 . To obtain one spectrum, averaging over 2000 scans was performed. The humidity of the air in the room during the measurements was controlled and was approximately 35%. The spectra of solutions were recorded in two identical cuvettes with different distances between the windows: 50.02 µm and 100.17 µm. The spectrum of the solution in the first cuvette was considered to be the background spectrum, and the spectrum of the same solution in the second cell was considered to be the spectrum of the sample. 

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