• M. A. MOROZOVA Peoples Friendship University of Russia (RUDN University), 6 Miklukho-Maklay Str.,117198, Moscow, Russia
  • A. M. KOLDINA Peoples Friendship University of Russia (RUDN University), 6 Miklukho-Maklay Str.,117198, Moscow, Russia
  • T. V. MAKSIMOVA Peoples Friendship University of Russia (RUDN University), 6 Miklukho-Maklay Str.,117198, Moscow, Russia
  • A. V. MARUKHLENKO Peoples Friendship University of Russia (RUDN University), 6 Miklukho-Maklay Str.,117198, Moscow, Russia
  • I. A. ZLATSKY Laboratory of Applied Biotechnologies, State Institute of Genetic and Regenerative Medicine NAMS of Ukraine, 67 Vyshgorodska Str., Kyiv 04114, Ukraine
  • A. V. SYROESHKIN Peoples Friendship University of Russia (RUDN University), 6 Miklukho-Maklay Str.,117198, Moscow, Russia


Objective: To investigate kinetic changes in the spectral characteristics by Fourier Transform Infrared spectroscopy (FTIR) of water-lactose complexes (SMC), derived during the manufacturing process of the drug, containing release-active forms of antibodies.

Methods: lactose monohydrate substance, saturated with release-active forms of affinity-purified polyclonal rabbit antibodies to recombinant human interferon-gamma (RA forms of Abs); tablets produced from this substance by direct compression after the addition of excipients (microcrystalline cellulose, magnesium stearate). Powdered and tableted placebo samples saturated with technologically processed water or phosphate-buffered saline, as well as with intact ethanol were used as control. Kinetic changes in SMC were studied using an Agilent Cary 630 FTIR spectrophotometer with a diamond ATR accessory (Agilent Technologies, USA). We used the method of X-ray fluorescence spectroscopy (EDX-7000 Shimadzu energy dispersive X-ray fluorescence spectrometer) to track changes in the fluorescence signal at certain wavelengths. The range of measured elements–11Na-92U.

Results: Control of some technological characteristics of the obtained active substance (moisture, flowability) and dosage form (mean mass, disintegration rate) was used as indirect indicators of quality, but they did not allow reliably distinguishing intact lactose from the saturated one. Long-period oscillations on FTIR spectra were characteristic for all types of samples; oscillations occur at approximately two-week intervals; S/N indices were more stable for samples of RA forms of Abs than for placebo samples. On some days, the substance saturated with RA forms of Abs significantly differed from the intact lactose powder. The kinetics of the X-ray fluorescence intensity at certain wavelengths indicates the possibility of a periodic cooperative trigger transition of the system. Reversible conformational transitions are observed for powders on the 30th and 130th days (Kα 3.313 keV). For tablets at Kα 3.313 keV and Kα 1.740 keV small changes were visualized on those days (100–110th day) when hysteresis phenomena were recorded in the IR spectra of these samples.

Conclusion: As a result, the evidence for a long-period dramatic conformational mobility of the water-lactose complex was obtained. Based on the data on the semiannual kinetics of IR spectra, a universal criterion for the identity of lactose powder saturated with RA forms of Abs was obtained. Also, it was confirmed that the lactose conformation state was changed by saturation with RA forms of Abs.

Keywords: Release-active forms of antibodies, Slow kinetic transitions, X-ray fluorescence spectroscopy, FTIR, Lactose monohydrate, Supramolecular complexes


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