According to the Federal Law «On Biomedical Cell Products», preclinical trials are an integral part of biomedical cell product (BCP) development. This article describes the basic principles of fulfilling requirements laid down in the Rules for conducting preclinical trials of BCPs. The main objective of preclinical trials is evaluation of efficacy, safety and biodistribution of cell products. Properly justified markers of biological activity must be used for reliable identification of BCP pharmacodynamic action in the host organism. BCP safety assessment must be comprehensive and include identification, characterization and quantitative evaluation of potential local and systemic toxicity, estimation of the onset of toxicity and possibility of its reduction, and the effect of a particular drug dose on the results of toxicity studies. The ultimate goal of preclinical toxicity studies is to obtain data sufficient for making a conclusion on the possibility of conducting clinical trials of BCP and determining associated risks. The key principles of preclinical trials design are a rational approach and justification of all decisions made during the study. The results of preclinical trials that were conducted in compliance with the law, can be included in the BCP dossier and considered during the product authorization by the expert institution of the Ministry of Health.

The article analyses main epidemiological parameters of tuberculosis (TB) in children and adolescents in Russia. It was demonstrated that the incidence rate in this population, though decreasing, is still high. The disease incidence in children and adolescents arising from contacts with people with active TB is 30 and 25 times higher than the overall incidence rate in children and adolescents in Russia, respectively. Analysis of TB lesions in a major proportion of children and adolescents shows multi-drug resistant strains of Mycobacteria tuberculosis (MTB), and this calls for revision of approaches to preventive actions for this population. The tuberculine Mantoux test is of limited value when it comes to diagnosing TB in adolescents. Screening in adolescents and children older than 7 years is performed using Diaskintest® containing two MBT-specific antigens. Specific TB prevention for newborns is performed by vaccination with BCG and BCG-M living vaccines. The product was modified by a 30 % reduction of the number of BCG living cells per 1 mg of vaccine in order to reduce postvaccinal complications largely associated with residual virulence of the BCG substrain. Since 2012 BCG vaccine lots have been standardized to contain from 500 thousand to 1 million viable BCG cells per BCG vaccine dose and from 375 thousand to 575 thousand viable BCG cells per BCG-M vaccine dose.

The article summarises methods used to control the quality of recombinant interferon alpha and beta (IFNs) products. It demonstrates that currently used methodological approaches generally ensure proper quality control of this group of medicines. However, it is necessary to formulate basic requirements for the pharmacopoeial general chapters on interferon alpha-2 and beta-1 products in order to standardize the control procedures and increase the efficacy and safety of recombinant IFNs products. These requirements should be harmonized with requirements of both the European Pharmacopeia and pharmacopoeias of the Eurasian Economic Union member-states, and should take account of the current trends in the Russian pharmaceutical industry.

The article describes a new method for measurement of monoclonal antibody Eculizumab concentration in human plasma in a range of 3-250 μg/ml. In this method we used an antibody Fab fragment, capable of binding to eculizumab specifically in human plasma. The method uses the biolayer interferometry for determination of Eculizumab concentration without additional tag in the analyzing substances. This method was comparatively validated along with the traditional ELISA. Comparative validation demonstrated that the biolayer interferometry based method has an advantage to the ELISA in such important statistical criteria as an analytical range, accuracy, precision, specificity and selectivity (matrix effect).

Identification and purity testing of biologicals as well as determination of their molecular mass are performed on the basis of comparison of test and reference samples by various physico-chemical methods, including Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) under reducing or non-reducing conditions. In order to assess the molecular weight of the test sample and reference sample components a calibration curve is drawn using protein markers. The article describes 4 experiments in which gel electrophoresis was used for separation of the most widely used sets of molecular weight markers produced by various manufacturers (Amersham™ LMW Calibration Kit for SDS Electrophoresis (Am), SDS PAGE Molecular Weight Standards, low range (BR), Unstained Protein Molecular Weight Marker (Th), BenchMark™ Protein Ladder (BM), Mark12™ Unstained Standard(M12) (total number of proteins - 45)). The aim of the study was to optimize mathematical processing of calibration curves generated for the above-mentioned sets of markers and to assess their interchangeability. SDS PAGE was performed under reducing conditions using Tris-Glycine gels. The real calculated ( M ) and nominal ( M₀) values (indicated in the set of markers instructions for use) of the protein markers molecular mass (MM) were compared using the MM deviation expressed in terms of a molecular mass unit (hereinafter - relative deviation). The estimated average MM relative deviation for sets of markers ranging from 10 to 100 kDa (Am, BR, Th sets), which was calculated using linear regression equations, was equal to about 10 % (8.3 - 12.0 %). In the case of the extended MM range of 10 - 220 kDa (BM, M12 sets) the average relative deviation was almost twice as high (16.5 - 20.3 %). A third-order polynomial regression equation was used to optimize mathematical processing methods used during calculation of the calibration function. This made it possible to reduce the MM relative deviation down to 2.3 - 3.5 % when using sets ranging from 10 to 100 kDa, and to 3.7 - 4.0 % when using the extended range sets, which is important for assessing similarity of different products.

Mycoplasmas are the main contaminants of cells cultures. They persist in cell cultures and can extensively affect host cell functions. Conducting experiments or producing protein in contaminated cultures are impractical. The aim of the study was to explore the possibility of quick detection of mycoplasma contamination of cell cultures and biotechnological products by a previously developed method which was modified to make it simpler and more affordable in Russia; and to assess the possibility of method validation for quality control. The authors chose the most applicable qPCR method of all qPCR methods currently used for mycoplasma detection, and modified it in the following way: expensive MGB-probes which cannot be synthesized in Russia were substituted by ordinary fluorescence probes. The reproducibility and sensitivity of the modified method were tested with M. hominis. The sensitivity of the test was equal to 10 mycoplasma gene copies per reaction. Comparison of the obtained results with regulatory requirements for mycoplasma detection showed that the proposed method complies with current official requirements and could be used as the main method for routine prompt cell culture testing for mycoplasma contamination.

The construction of highly sensitive new-generation diagnostic test systems requires the use of modern innovative technologies that are capable of detecting infectious agents at the molecular level by identifying specific antibodies to individual immunoreactive epitopes. The development of such immunoassays is based on molecular mapping of a target antigen using a library of short overlapping peptides aimed at identification of both epitopes specific for a particular pathogen, and those common to the group of infectious agents. The article describes new approaches that are based on the results of mapping of the Omptin group model protein that could be critical for improving retrospective laboratory diagnosis of infectious diseases caused by bacteria of the Enterobacteriaceae family. The article discusses the prospects for assessing the applicability of the peptide ELISA experimental version, designated as p6/24-Omptin-TIFA, for the detection of antibodies to diagnostically significant Omptin epitopes in the serum of people who have had enterobacterial infections, in order to introduce it into healthcare practices.

Analysis of methodological approaches to preclinical evaluation of sensitizing activity of horse serum products shows that the anaphylactogenicity scale recommended in national regulatory documents is imperfect as it is not capable of fully predicting the safety of the clinical use of products. The article demonstrates feasibility of another methodological approach to preclinical evaluation of sensitizing activity of heterologous serum products which is based on calculation of a sensitizing dose inducing anaphylactic reactions in 50 % of guinea pigs. This approach will make it possible to more effectively predict side effects of already authorized heterologous immunoglobulins and drugs currently under development, and to select a clinical use strategy on a case-by-case basis.