Biopreparaty Profilaktika diagnostika lechenie最新文献

{"title":"Study of a combined biological product with antibacterial and probiotic activity","authors":"V. A. Neschislyaev, E. G. Shilova, A. M. Nikolaeva, E. V. Orlova","doi":"10.30895/2221-996x-2023-23-3-1-422-430","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-422-430","url":null,"abstract":"Scientific relevance . The issue of antimicrobial resistance has acquired global significance, urging the search for and use of alternative antibacterial agents to treat infectious diseases. In particular, this situation prompts wider use of biotechnology-derived medicinal products based on bacteriophages and probiotics, including those of the metabolite type. The bacteriotropic (antimicrobial and probiotic) properties of pharmaceutical compositions based on bacteriophages and metabolites of probiotic bacteria suggest a qualitatively new combined antibacterial effect, as well as effectiveness against microorganisms resistant to antibiotics. Aim . The authors aimed to study the properties of a combined biological product with antibacterial and probiotic effects. Materials and methods . The study focused on a combined biological product consisting of a probiotic agent based on Lactobacillus exometabolites and a complex bacteriophage. The antimicrobial activity evaluation involved the Appelmans method, a disc diffusion method using lawn cultures homologous ( Staphylococcus spp., Pseudomonas aeruginosa , Escherichia coli , Proteus spp., Enterococcus spp., Salmonella spp., and Shigella spp.) and non-homologous ( Acinetobacter baumannii, Klebsiella pneumoniae , and Yersinia enterocolitica ) to the complex bacteriophage, and a bioluminescence inhibition test using the genetically modified indicator strain E. coli lum+. The study used bacterial strains isolated from various clinical samples and biotopes in bacteriological laboratories of healthcare institutions in the Perm Territory in 2019–2022. The probiotic activity was assessed by the stimulating effect on model microorganisms of the normal flora. Results . The Appelmans method showed that the combined biological product had high antimicrobial activity against microorganisms homologous to the complex bacteriophage. The titres calculated for the combined biological product and its complex bacteriophage component were comparable and amounted to 10 –6.6±0.01 and 10 –6.9±0.01 , respectively. The disc diffusion method demonstrated that the combined biological product had a more pronounced antibacterial effect on all tested strains than its individual components. The optical density values obtained with the combined biological product were 1.5 and 2 times higher than the values observed with control samples in Bifidobacterium bifidum 1 and Lactobacillus plantarum 8P-А3 cultures, respectively, which demonstrated the stimulating effect of the product on the normal flora. The study results suggest the compatibility of the phage and probiotic components of the combined biological product, as well as its high antimicrobial activity. Conclusions . The novel combined biological product has high specific and non-specific antimicrobial activity, which consists in the inhibition of pathogenic bacteria growth without affecting the normal flora. The combined biological product broadens the range of medicinal products ","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"53 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"134944965","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Probiotic activity of <i>Bacillus subtilis</i> metabolites in experimentally induced dysbiosis in mice","authors":"S. А. Lazarev, N. O. Vartanova, A. V. Poddubikov, N. A. Mikhailova","doi":"10.30895/2221-996x-2023-23-445","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-445","url":null,"abstract":"Scientific relevance . A promising option for dysbiosis correction is the use of metabiotics, products based on metabolites of probiotic microorganisms. During fermentation, Bacillus subtilis bacteria (strains 3H and 1719) produce metabolites that exhibit probiotic properties in vitro . These observations in vitro motivate an in vivo investigation of B. subtilis metabolite effects on colonic mucosal microbiota in mice in experimentally induced dysbiosis and an assessment of the potential of B. subtilis metabolites as metabiotics. Aim . The authors aimed to compare the probiotic activity of B. subtilis 3H and B. subtilis 1719 metabolites and a commercial metabiotic in antibiotic-induced dysbiosis in mice. Materials and methods . The authors induced experimental dysbiosis in BALB/c mice weighing 18–20 g by intraperitoneal injection of gentamicin. For subsequent correction, the test groups received sorbent-bound B. subtilis metabolites, and the comparison group received a commercial metabiotic containing B. subtilis metabolites (VKPM B-2335(3)3) via intragastric injection for 21 days. The quantitative and qualitative analysis of colonic mucosal microbiota included microbial culturing and colony identification by MALDI-TOF mass spectrometry. Results . Antibiotic-induced colonic dysbiosis in mice manifested itself as a decrease in the dominant microbiota and an increase in opportunistic pathogens. After 7 days of metabolite administration, the Lactobacillus population returned to normal in all treatment groups. The mice that received B. subtilis 3H metabolites showed the best results: their Lactobacillus spp. composition corresponded to that of intact animals. The content of Lac+ Escherichia coli returned to 100% in all treatment groups. After 21 days of metabolite administration, the authors observed the elimination of bacteria ( Rodentibacter spp., Aerococcus spp.) and fungi ( Trichosporon spp., Kazachstania spp.) in the B. subtilis 3H group; Trichosporon spp. (no effect on Kazachstania spp.) in the B. subtilis 1719 group; and Enterococcus spp., Kazachstania spp., and Trichosporon spp. (no effect on Rodentibacter spp. and Aerococcus spp.) in the commercial metabiotic group. Conclusions . Metabolites of B. subtilis strains 3H and 1719 help to restore the diversity and abundance of colonic microbiota in antibiotic-induced dysbiosis. The differences observed in microbiota re-establishment in the treatment groups indicate that there is interstrain variability in the probiotic activity of B. subtilis metabolites.","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"23 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135814328","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Determination of optimum nanofiltration conditions for the manufacturing process of human normal immunoglobulin G for intravenous administration","authors":"N. V. Zubkova, A. M. Nikolaeva, A. V. Ivanov, O. V. Beliakova, M. V. Razumikhin, N. V. Vinokurova, I. S. Efimova, T. I. Smolyanova, E. I. Sakanyan","doi":"10.30895/2221-996x-2023-23-3-1-400-410","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-400-410","url":null,"abstract":"Scientific relevance . Medicinal products based on immunoglobulin class G (IgG) from human plasma are widely used in clinical practice to treat bacterial and viral infections, primary and secondary immunodeficiencies, and autoimmune diseases. Nanofiltration is a way to mitigate the risk of in-process contamination of raw materials with various pathogens, including viruses. Therefore, it is relevant to investigate the development and implementation of additional viral inactivation and/or elimination steps. Aim . This study aimed to develop and validate optimum nanofiltration conditions and to scale up the nanofiltration step for the manufacturing of human IgG for intravenous administration. Materials and methods . The study used a solution of IgG from plasma fractions II and III. The authors paired nanofilters manufactured by Planova 20N and BioEx (Asahi Kasei, Japan), Viresolve Pro (Merck Millipore, USA), Virosart HC and HF (Sartorius, Germany), and Pegasus SV4 and Prime (Pall, USA) with Sartopore polyethersulphone prefilters by Sartorius (Germany), Virosart MAX polyamide prefilters by Sartorius (Germany), and EKX-P regenerated cellulose prefilters by Pall (Germany). Virus reduction validation studies were performed with model viruses (human immunodeficiency virus type 1, porcine transmissible gastroenteritis virus, porcine parvovirus, murine encephalomyocarditis virus, and bovine viral diarrhoea virus) in the laboratories of the N.F. Gamaleya centre. The sample data analysis involved calculating mean values with 95% confidence intervals. Results . For all the selected combinations of prefilters and filters, the maximum nanofiltration throughput depended on the IgG concentration in the test solution. With the combination of an EKX-P filter with a Pegasus SV4 nanofilter, the maximum throughput and the IgG yield reached 6300 g/m 2 and 95%, respectively. When combined with a Planova 20N nanofilter, EKX-P and Sartopore (polyethersulphone) filters provided a maximum throughput of up to 2980 g/m 2 and an IgG yield of almost 100%, provided that the test solution had an IgG concentration of 10 g/L. With different filter combinations, virus reduction levels ranged from 4.00±0.05 to 4.75±0.04 log 10 for human immunodeficiency virus type 1, from 4.30±0.04 to 4.55±0.06 log 10 for porcine transmissible gastroenteritis virus, from 5.38±0.08 log10 to 5.57±0.04 log 10 for murine encephalomyocarditis virus, 5.12±0.10 log 10 to 5.25±0.08 log 10 for porcine parvovirus, and exceeded 5.00 log 10 for bovine viral diarrhoea virus. The virus reduction levels achieved were not statistically associated with prefilter brands. Conclusions . The study demonstrated that nanofiltration was effective at removing viruses with various virion sizes and physicochemical characteristics, including viruses as small as parvovirus B19. The levels of virus reduction exceeded 4 log 10 and met the acceptance criteria. The laboratory-scale nanofiltration parameters and the corresponding fil","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135487068","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of quality attributes of a human albumin preparation with a modified stabilising composition","authors":"M. V. Tomilin, T. V. Korotkova, P. A. Loginov","doi":"10.30895/2221-996x-2023-23-3-1-411-421","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-411-421","url":null,"abstract":"Scientific relevance . The national and international human albumin preparations registered in the Russian Federation mainly differ in their excipient compositions. While all the international preparations of human albumin contain a mixture of sodium caprylate and N-acetyl-DL-tryptophan, the Russian ones contain only sodium caprylate. However, albumin stabilisation with sodium caprylate at high concentrations affects the ligand-binding properties of albumin. For this reason, as well as to achieve storage stability not only at temperatures of 2 °C to 8 °C but also at room temperature, most international manufacturers have reduced the sodium caprylate content in albumin preparations and added N-acetyl-DL-tryptophan. This demonstrates the relevance of studying the quality of a new Russian human albumin preparation with a modified stabilising composition, including both sodium caprylate and N-acetyl-DL-tryptophan. Aim . The study aimed at comparing several quality attributes of the human albumin preparation with a modified stabilising composition with those of imported human albumin preparations. Materials and methods . The human albumin preparation with a modified stabilising composition was manufactured by fractionation from donor plasma meeting the requirements of monograph FS.3.3.2.0001.19 of the State Pharmacopoeia of the Russian Federation edition XIV. The quality control was in line with the monograph on human albumin (FS.3.3.2.0006.18), and statistical analysis was conducted in Microsoft Excel in accordance with the general chapter on statistical analysis (OFS.1.1.0013.15). Results . The study preparation complied with the requirements specified in monograph FS.3.3.2.0006.18. All the manufactured batches were clear, thermostable, sterile, and non-pyrogenic. The prekallikrein activator levels were low (below 1 IU/mL). The aluminium content varied from 30.36±10.39 µg/L to 50.22±6.94 µg/L. The study preparation contained sodium ions at a concentration from 127.44±10.46 mmol/L to 145.59±7.32 mmol/L and less than 0.01 mmol/g of potassium ions. The osmolarity exceeded 240 mOsm/L. The content of α- and β-globulins ranged from 1.79±0.06% to 2.24±0.20%. The study preparation had a pH level of 6.9 to 7.2. The concentrations of polymers and aggregates did not exceed 0.5%. Conclusions . The quality attributes studied suggest that the human albumin preparation with a modified stabilising composition is comparable to its international counterparts and that it meets Russian and European pharmacopoeial standards.","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"49 2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135786750","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development, certification, and use of a pharmacopoeial standard for the content of immunoglobulin A in human immunoglobulins for parenteral administration","authors":"A. V. Nechaev, E. Yu. Kudasheva, E. L. Postnova, R. A. Volkova, O. V. Fadeikina, I. V. Borisevich, A. A. Movsesyants","doi":"10.30895/2221-996x-2023-23-3-1-443-451","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-443-451","url":null,"abstract":"Scientific relevance . The immunoglobulin A (IgA) impurity content in parenteral human immunoglobulins should be determined in accordance with the State Pharmacopoeia of the Russian Federation by kinetic nephelometry, radial immunodiffusion, or enzyme immunoassay (ELISA) with a reference standard. The International Standard (IS) for the content of IgA is certified using gravimetry and radial immunodiffusion. However, neither of the existing standards for the content of IgA in human immunoglobulins is currently certified using all three compendial methods. This prevents analysts from comparing test results obtained by different methods and may lead to an underestimation of the IgA content in human immunoglobulins. Aim . This study aimed to determine the procedure for the development, certification, and use of a pharmacopoeial reference standard (RS) for the content of IgA in human immunoglobulins. Materials and methods . The authors studied candidate RSs for the IgA content derived from human plasma for fractionation. The IgA content determination involved kinetic nephelometry, radial immunodiffusion, and ELISA, as well as commercial test kits and the IS. The authors quantified the IgA impurity in samples of commercial human immunoglobulins from various manufacturers. The data analysis involved descriptive statistics and variance analysis using Microsoft Excel and Statistica 10. Results . The authors established a pharmacopoeial standard with a certified IgA content of 1.98 mg/mL (expanded uncertainty, 0.44 mg/mL; coverage coefficient, k =2; confidence level, 95%) for IgA impurity quantification in human immunoglobulins by radial immunodiffusion and ELISA and that of 1.31–2.64 mg/mL (expanded uncertainty, 0.67 mg/mL; coverage ratio, k =3; confidence level, 99%) for intralaboratory quality control of IgA impurity quantification by kinetic nephelometry, radial immunodiffusion, and ELISA. Conclusions . The pharmacopoeial standard developed in the study has been included in the register of standards of the State Pharmacopoeia of the Russian Federation as the Reference Standard for the Content of Immunoglobulin Class A (IgA) (Registry No. 3.1.00454). The pharmacopoeial standard is intended for the standardisation of analytical methods for the-determination of the IgA impurity content in parenteral human immunoglobulins.","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"22 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135786905","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Effects of pertussis toxin and &lt;i&gt;Bordetella pertussis&lt;/i&gt; lipo-oligosaccharide on the specific toxicity and potency of whole-cell pertussis vaccines","authors":"I. A. Alekseeva, I. V. Ibragimkhalilova, D. N. Lepekhova","doi":"10.30895/2221-996x-2023-23-3-333-347","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-333-347","url":null,"abstract":"Scientific relevance. The content of Bordetella pertussis lipo-oligosaccharide (LOS) and the residual levels of active pertussis toxin (PT) are generally accepted to be the primary factors that determine the reactogenicity of whole-cell pertussis vaccines. To improve the quality of whole-cell pertussis vaccines, it is both relevant and necessary to study the relationship between the toxicity of B . pertussis bacterial cell components and the main quality parameters of these vaccines, including their potency and specific toxicity, as termed in the WHO recommendations and the European Pharmacopoeia. Aim . This study aimed to analyse the effects of B . pertussis LOS and residual active PT on the specific toxicity and potency of adsorbed diphtheria, tetanus, and whole-cell pertussis (DTwP) vaccines. Materials and methods . The authors tested 57 commercial batches of adsorbed DTwP vaccines for compliance with the regulatory standards and product specification files. Vaccine batches that failed specific toxicity tests formed Group 1, and the other batches were designated as Group 2. The potency was tested in F1 CBA/Ca×C57BL/6J hybrid mice with experimentally induced meningoencephalitis that were immunised with DTwP and reference vaccines. The authors assessed the specific toxicity of DTwP vaccines by changes in body weight following intraperitoneal administration. The toxic activity was assessed indirectly by changes in body weight in the first 16–24 h ( B . pertussis LOS) and on day 7 (PT) after dosing. The authors used Spearman’s rank correlation coefficient to measure the strength of correlation between the toxic activity of vaccine components and the specific toxicity and potency of the vaccine, which were established using the same vaccine batches. Results . The authors measured the toxic activity of LOS and residual active PT in the vaccine batches studied. The correlation coefficients between the specific toxicity and potency of the vaccines and the toxic activity of LOS were 0.113 ( p >0.05) and 0.049 ( p >0.05), respectively. Similarly, the correlation coefficients between the specific toxicity and potency of the vaccines and the toxic activity of PT accounted for 0.595 ( p <0.01) and –0.534 ( p <0.01), respectively. Conclusions . The authors studied the toxic activity of B. pertussis LOS and residual active PT in whole-cell pertussis vaccines and found an inverse correlation between the potency of the vaccines and the toxic activity of residual active PT. The study demonstrated that the specific toxicity test for whole-cell pertussis vaccines fails to detect and quantify B. pertussis LOS in the samples. The authors advise to determine the content of LOS in the B. pertussis strains intended for the production of whole-cell pertussis vaccines, which is not yet an accepted practice in the Russian Federation.","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"55 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135786376","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Biopharmaceutical properties of polyvalent bacteriophage capsules","authors":"A. M. Nikolaeva, N. A. Kovyazina, E. V. Funkner, A. N. Krasilnikova, K. A. Lysko","doi":"10.30895/2221-996x-2023-23-3-1-379-388","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-379-388","url":null,"abstract":"Scientific relevance. Rational phage therapy is a highly effective way to combat bacterial infections, especially in conditions of steadily increasing antibiotic resistance. Most bacteriophage preparations are formulated as oral and topical solutions. However, oral administration of liquid phage preparations results in significant inactivation in the stomach. To shield active ingredients from the acidic environment, Sextaphag ® Pyobacteriophage, polyvalent, has been formulated into capsules. Aim . This study evaluated the polyvalent bacteriophage preparation in capsules in terms of its potency and biopharmaceutical properties. Materials and methods. The study compared the polyvalent bacteriophage preparation formulated as capsules with the polyvalent bacteriophage preparation formulated as a solution. The potency was evaluated by the Appelmans method, and phage particles were quantified by the Gratia method. To evaluate the acid-neutralising capacity, the authors placed test samples of the bacteriophage preparation in 0.1 M hydrochloric acid and analysed their potency by the Appelmans method. Chinchilla rabbits were used to analyse anti-phage immune responses, and their antibody levels were measured using an enzyme immunoassay test system developed by the authors. The pharmacokinetic parameters were studied in outbred white mice after oral dosing. Results. The polyvalent bacteriophage preparation exhibited high lytic activity towards Escherichia coli , Klebsiella pneumoniae , Proteus mirabilis , Proteus vulgaris , Pseudomonas aegidinosa , Staphylococcus aureus, and Streptococcus pneumoniae , which accounted for a dilution factor of 10 -6 . Following oral administration of the polyvalent bacteriophage preparation in capsules to mice, the level of absorption was 3.1–3.7 times higher than that observed with the solution. Repeated oral administration of therapeutic doses did not induce anti-phage antibodies in rabbits. The stability study showed that the polyvalent bacteriophage preparation retained high lytic activity for 18 months. Conclusions. According to the study results, the polyvalent bacteriophage preparation in capsules exerts significant antibacterial activity against the studied microorganisms, has a high level of absorption, retains its lytic activity for a long time, and does not induce anti-phage antibodies after oral dosing, which confirms its safety and efficacy.","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"33 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135786751","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Estimation of measurement uncertainty for the determination of loss on drying of biologicals","authors":"O. V. Fadeikina, A. A. Voropaev, D. S. Davydov, R. A. Volkova","doi":"10.30895/2221-996x-2023-23-3-1-452-462","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-452-462","url":null,"abstract":"Scientific relevance . GOST ISO/IEC 17025-2019 requires testing laboratories to evaluate the measurement uncertainty of their results. Estimating the uncertainty of analytical methods intended for biologicals is a challenging task that requires time, effort, and a special approach. Measurement uncertainty estimation is of particular interest in the case of measuring loss on drying (LOD) for biologicals, since LOD testing procedures involve analysing measurements of a physical value, i.e. mass. Aim . This study aimed to estimate the measurement uncertainty of LOD determination in biological medicinal products. Materials and methods . The study examined a powdered active substance intended for a Bifidobacterium product (test sample). The authors conducted the LOD test in accordance with the State Pharmacopoeia of the Russian Federation (OFS.1.2.1.0010.15). Statistical processing of the results was performed using Microsoft Excel. To estimate the measurement uncertainty, the authors employed the bottom-up approach or used the standard deviation from testing results. Results . The authors identified the uncertainty components that affected the LOD determination results. When calculated using the bottom-up approach, the expanded uncertainty was 0.34% (coverage factor, k =2; approximate confidence level, 95%). In particular, the largest contributor to the expanded uncertainty was the uncertainty of measuring the mass of weighing bottles containing dried test samples (0.147%), whereas the smallest contributor was the uncertainty of weighing empty bottles (0.003%). When calculated using the standard deviation, the uncertainty of two parallel measurements amounted to 0.32%. Conclusions . Both approaches to calculating LOD measurement uncertainty yield comparable results. According to the uncertainty budget analysis, the uncertainty of measuring the mass of weighing bottles with dried test samples is the major contributor to the test result. For this reason, the conditions of sample preparation should be carefully controlled. The study results confirm that the LOD measurement uncertainty can be calculated using the standard deviation. Testing laboratory teams may benefit from the methods for identifying the factors influencing LOD test results and the methods for calculating the uncertainty of measurement described in this study.","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"242 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135786752","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"On assessing the viral safety of individual units of the substance \"Human plasma for fractionation\" by nucleic acid amplification","authors":"E. V. Elbert, V. V. Nozhko, R. A. Volkova, A. A. Movsesyants, V. A. Merkulov, V. V. Kosenko","doi":"10.30895/2221-996x-2023-23-3-1-463-473","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-463-473","url":null,"abstract":"Scientific relevance . The absence of blood-borne viruses in human plasma-derived medicinal products must be ensured by the control of raw materials and the manufacturing process. Aim . This study aimed to analyse system suitability criteria for analytical procedures to assess the viral safety of individual units of the substance \"Human plasma for fractionation\" in terms of the content of nucleic acids of blood-borne viruses, considering the requirements of the European Pharmacopoeia. Materials and methods . The authors analysed individual units of the substance \"Human plasma for fractionation\" (hereinafter, plasma). The study used the International Standards (ISs) for human immunodeficiency virus RNA, hepatitis A virus (HAV) RNA, hepatitis C virus (HCV) RNA, hepatitis B virus (HBV) DNA, and parvovirus B19 DNA, as well as nucleic acid detection kits for these viruses based on polymerase chain reaction (PCR). Results . HCV RNA was not detected in any of the eight plasma samples studied, and parvovirus B19 DNA was detected in one of the samples at a concentration not exceeding 10 4 IU/mL. Three tests with the corresponding ISs showed that the studied reagent kits detected HCV RNA at a concentration of 10 2 IU/mL and parvovirus B19 DNA (M1 genotype) at a concentration of 10 4 IU/mL. In additional tests that were conducted in two samples considering the requirements of the European Pharmacopoeia for the detection of HCV RNA and parvovirus B19 DNA, a new batch of reagent kit I detected the HCV RNA IS at a concentration of 102 IU/mL only in one of three replicates, which did not correspond to the claimed sensitivity of the reagent kit. HCV RNA was not detected in either replicate in one of two plasma samples spiked with the HCV RNA IS at concentrations of 10 2 and 10 3 IU/mL, possibly because of plasma inhibitory properties. The sensitivity of the reagent kits to parvovirus B19 DNA corresponded to the label claims; the study did not show any inhibitory properties of the plasma samples. Conclusions . Polymerase chain reaction testing of the viral safety of plasma intended for manufacturing medicinal products should include control samples calibrated in IU/mL. Further research and appropriate pharmacopoeial reference materials are needed to set system suitability criteria for analytical procedures using such control samples.","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"69 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135786749","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Pollen allergen products: current standardisation issues","authors":"E. I. Sakanyan, M. A. Yasnaya, V. F. Vul, R. A. Bubenchikov, N. V. Vinokurova, E. S. Yurtaeva","doi":"10.30895/2221-996x-2023-23-3-1-367-378","DOIUrl":"https://doi.org/10.30895/2221-996x-2023-23-3-1-367-378","url":null,"abstract":"Scientific relevance . Pollen allergen medicines are in high demand, and their therapeutic benefits directly correlate with their standardisation. Better diagnosis and treatment of allergic diseases require state-of-the-art procedures for assessing the allergenic activity of pollen allergen products using reference standards and physicochemical testing methods. Aim . The study aimed at developing methodological approaches to the standardisation of pollen allergen products in order to shift to measuring their potency in allergenic activity units (AAU) and bring their quality in line with the requirements of the State Pharmacopoeia of the Russian Federation. Materials and methods . The study used pollen allergen reference standards by Microgen, the WHO International Standard for timothy grass ( Phleum pratense ) pollen extract, a gel filtration standard kit of molecular weight markers ranging from 1.35 to 670 kDa, bovine serum albumin, serum samples with specific IgE obtained from donors sensitised to the study pollen allergens, labelled anti-human IgE antibodies, and reference standards for determining residual volatile solvents by gas chromatography. The identification of in-house reference standards for the potency of pollen allergens involved Western blotting (for allergenic components). The total protein content was determined by Bradford’s assay. In addition, the authors used high-performance liquid chromatography to study protein fractions and gas–liquid chromatography to determine the content of residual organic solvents. Results . To substitute the existing method of non-specific characterisation of allergenic activity in protein nitrogen units (PNU), the authors developed and tested a new method to control allergenic activity in allergenic activity units (AAU) based on an in vitro competitive enzyme-linked immunosorbent assay (ELISA) procedure developed and validated in this study. Furthermore, the authors developed and certified 15 primary in-house reference standards with allergenic activity established in AAU/mL using skin tests in vivo . The experimental data were analysed to standardise the allergenic activity of the pollen allergens manufactured by Microgen. The authors developed physicochemical methods for the certification of in-house reference standards and validated these methods in accordance with the State Pharmacopoeia of the Russian Federation. The study involved selecting chromatographic separation conditions for residual organic solvents (acetone and diethyl ether) and establishing system suitability criteria for the chromatographic system. The allergenic activity of secondary in-house reference standards was certified against that of primary in-house reference standards using competitive ELISA. Thus, the authors managed to shift to the standardisation of pollen allergen products in vitro . Conclusions . The authors developed their competitive ELISA-based method to standardise pollen allergen products by comparing the inhi","PeriodicalId":32767,"journal":{"name":"Biopreparaty Profilaktika diagnostika lechenie","volume":"14 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-09-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135786904","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}