Biotechnological products manufactured by recombinant DNA technology are widely used nowadays. According to the national and  international requirements the amount of residual host cell DNA in  such products should not exceed 10 ng per dose. However, for  products intended for frequent or long-term use, this amount must  not exceed 100 pg per dose. This article describes methods most  frequently used for quantification of residual host cell DNA in  biological active substances contained in biotechnological products:  molecular hybridization with biotin- or digoxigenin-labelled DNA- probes (semiquantitative method), Threshold system, real-time PCR, method based on the use of a fluorescent reagent (assays). If  a method based on the use of a fluorescent reagent or real-time PCR are used to replace the current procedure, it is necessary to  demonstrate their validity, e.g. by comparing the results of residual DNA quantification obtained by the two methods — the new one and the current one. The article dwells upon the advantages and  disadvantages of the methods and potential sources of uncertainty.  It highlights the importance of using appropriately certified reference standards and retention samples. The biological active substances  included into the State Register of Medicinal Products conform to the international requirements in terms of the amount of residual host cell DNA.

Addition of adjuvants is a generally recognized method of enhancing the immunogenicity of vaccines. There is a large variety of  adjuvants, therefore the choice of an adjuvant is based on the  comparison of adjuvants efficacy in animal models, as well as  assessment of their safety and tolerability. There are many different  groups of substances that may have adjuvant properties, but  relatively little attention is paid to carbohydrate-based adjuvants, despite the fact that they are compatible with live vector  vaccines, safe, well tolerated, and their production is not laborious.  Thanks to these advantages the use of these particular adjuvants in  combination with any type of vaccine, including vector vaccines and  DNA vaccines, may be most appropriate and promising. This review  examines various types of carbohydrate adjuvants and the  mechanisms that determine their efficacy. 

Medicinal products based on pollen allergen extracts are widely used for diagnosis and therapy of pollinosis. According to the  requirements of the State Pharmacopoeia of the Russian Federation,  13th ed., the allergenic potency of such products is assessed against certified reference materials whose biological activity was confirmed in vivo in subjects sensitized to particular tested allergens. The finished dosage form of a candidate In-House Reference Material (IHRM) of timothy pollen allergen extract and the corresponding  allergen pollen extract (intermediate) were characterized in terms of  protein profile and allergenic composition using SDS-PAGE, gel- filtration HPLC, and western blotting. The test samples were  compared to finished pharmaceutical products (FPP) and pollen  allergen extracts produced at different dates and made from pollen  harvested at different dates, as well as to the WHO International  Standard (IS) of Timothy Pollen Extract (NIBSC code: 82/520). SDS- PAGE and gelfiltration HPLC showed that the candidate IHRM protein  profile was comparable in terms of major protein fractions to those  of all the FPP batches and pollen allergen extracts produced at  different dates. HPLC confirmed the comparability of the protein  profiles of the candidate IHRM and the IS (NIBSC code: 82/520), but showed minor  variations in the ratio of the main protein fractions.  Western blotting confirmed the presence of the main allergenic components with relative molecular masses ranging from 50 to 60  kDa and from 27 to 35 kDa. The composition of specific allergenic  components of timothy pollen allergen products manufactured by  JSC «SIC «Microgen» was identical with that of the WHO IS of  Timothy Pollen Extract in terms of the main protein and allergenic  components. 

Water for injections is one of the most popular diluents used for preparation of parenteral dosage forms. The European  Pharmacopoeia recommends two methods for the determination of  subvisible particulate matter: Light Obscuration Particle Count Test  and Microscopic Particle Count Test. The Russian Pharmacopoeia,  13th ed., additionally allows for the use of the Coulter principle  (Electrical Sensing Zone method). Thus, a procedure had to be developed for subvisible particles determination in water for  injections based on the Coulter principle (hereinafter — procedure).  The article presents the results of development and validation of the  procedure, i.e. the characteristics of accuracy, dilutional linearity,  ruggedness in terms of the time factor, and repeatability for particles more than 10 μm in size. The results of subvisible particles  determination obtained with the help of the developed procedure based on the Coulter principle were compared to the  results obtained with the help of the light obscuration particle count  test. The accuracy of the developed procedure was supported by the  statistical insignificance of the differences between the obtained  results. The values of ruggedness in terms of the time factor (NMT  14 %) and repeatability (NMT 15 %) did not exceed the established  acceptance criterion which is equal to the acceptable limit of the  instrument error for particle count in the tested samples (20 %). The  dilutional linearity of the procedure was demonstrated  (coefficient of determination R2 = 0.999). The results obtained  during the validation studies support the possibility of using the  Coulter principle for the assessment of subvisible particles in water  for injections.

The production of biological medicinal products used for prophylactic, diagnostic and therapeutic purposes includes the use of inherently  variable biological processes and materials. In order to ensure the  quality of the finished biological product it is necessary to determine  the source, nature and fitness for use of the starting materials, including reagents, culture media, buffer solutions, sera,  and enzymes. The article summarises literature data on the  structure, properties and mode of action of the animal-derived  reagent trypsin — proteolytic enzyme widely used in the production  of biological medicinal products. The article dwells upon the sources  of trypsin, methods of its production, requirements for trypsin  products of different purity grades intended for human and  veterinary use as well as for use as reagents in the production of  vaccines, advanced therapy medicinal products and genetically engineered products. The article describes the role that  trypsin plays in the human and animal intestinal digestive enzyme  systems, in dissociation of cells in cultures in the process of cell  passaging, in the mechanism of proteolitic activation and inactivation of a wide range of viruses, and in the examination of proteins  primary structure.

The article analyses the efficacy of vaccination and its influence on the nature of tuberculosis process in children perinatally exposed to  HIV. The study analysed hospital records, hospital discharge  summaries and outpatient medical records of children under 14 with  tuberculosis: there were 109 HIV-infected children and 97 children  perinatally exposed to HIV (but not HIV-infected). The postvaccinal  immunity status was assessed by the presence and size of the  vaccination scar and the response to the Mantoux test. The clinical  efficacy of the BCG vaccination was assessed by the severity of  tuberculosis in vaccinated and non-vaccinated children. It was shown that immunization with BCG vaccine (BCG-M) was safe and  efficacious in children born to HIV-infected women but not infected  with HIV. At the same time the vaccine did not demonstrate  sufficient immunological and clinical efficacy in HIV-infected children: the response to the Mantoux test, 2 TU, was positive only in one  third of all vaccinated children; there was no statistically significant  difference in the frequency of disseminated processes in vaccinated children as compared to the non-vaccinated children  perinatally exposed to HIV. A conclusion was made that children born to HIV-infected mothers but not infected with HIV must be  vaccinated during the neonatal period. The vaccination of HIV- infected children is not advisable due to its low clinical efficacy and  the risk of development of disseminated complications, for instance,  the generalized BCG infection in HIV-infected children may develop over a period of 3 years after vaccination.

The efficacy of influenza vaccines has been a matter of considerable debate ever since the development of the first influenza vaccine. The efficacy of currently used influenza vaccines depends on many  factors, including the strain composition, the degree of homology  between the produced and epidemic influenza viruses, the  vaccination coverage, and many other factors. Assessment of quality, i.e. determination of compliance of the product’s quality  characteristics with the specification requirements, and assessment  of risks associated with the use of the product were considered only  in the context of general requirements for the quality of biologicals.  The article summarises the results of analysis of batch to batch  consistency of the influenza inactivated polymer-subunit vaccine. The study included a retrospective assessment of the data  obtained during the product release control and testing performed by an accredited testing centre as part of mandatory certification. The  data obtained may be used to improve the production method and  the system of statistical management of the production process, as  well as to assess the risks accompanying the production of influenza vaccine at each of the stages.