This article presents the results of a study of the effectiveness of anti-rabies and anti-chlamydia antibodies conjugated with fluorescein, freeze-dried. Immunobiological products lyophilization is one of the most reliable ways to preserve their physicochemical and biological properties, since this is a soft drying method, in which the dried product is frozen, then it is placed in a vacuum chamber, where the solvent is sublimated. Lyophilization allows you to get drugs without losing their shape, structural integrity and more importantly, without losing their biological activity. The study aimed to standardize methods for the purification of FITC conjugates, as well as reagents used in the production and drying of FITC conjugates. We used conjugates of anti-rabies and anti-chlamydia antibodies labeled with fluoresceinisothiocyanate (FITZ) obtained by hyperimmunization of sheep and rams using an immunostimulant. We tested the ability of various concentrations of dextran T-70 to retain the specific activity of fluorescent conjugates of anti-rabies and anti-chlamydia antibodies when added dextran T-70 to them before the lyophilization. To determine the optimal amount of T-70 dextran filler, prepared 1%, 2%, 3%, 4%, and 5% solutions in 10mm phosphate-salt solution. We found that the effectiveness of dextran T-70 as a filler substance in the lyophilization of FITC conjugates depends on its concentration. The optimal stabilizing vehicle for lyophilization of both fluorescent conjugates is 2-3% concentration of dextran T-70 in 10 mM phosphate-buffered saline. Analysis of the results of the quality of FITZ-conjugates allows us to conclude about the high quality of biological products prepared during freeze-drying of the material in the developed temperature regime using dextran T-70 as a filler substance. As a result of the studies, the technology for the preparation of lyophilized FITC conjugates of anti-rabies and anti-chlamydia antibodies for diagnostic test systems was optimized. The specific activity and stability of the physicochemical characteristics of the preparations after lyophilization and during subsequent storage for two years were observed.
Abstract. This article summarizes the results of a study of the cultural properties of the swine reproductive and respiratory syndrome virus isolate in transferable cultures of MA-104, PK-15, MARC-145 and Vero cells. The purpose of our research was to determine the sensitivity of cell culture lines to the swine reproductive and respiratory syndrome virus isolate, which is necessary for obtaining a highly active antigen as the main component of diagnostic and vaccine biologics. An isolate of the virus was isolated from a Piglet in the LPH of the Moscow region of the Kolomenskoye district. The isolated isolate of an infectious disease of pigs by the method of molecular biological analysis is characterized in the established order for this pathogen. Novelty. The possibility of reproduction in MARC-145 cell culture has been demonstrated. Results. It was shown that during reproduction of the virus isolate in cell culture for 96 ± 6 hours at a dose of 0.1 TCD50 / cell infection, an antigen with high biological activity was obtained. Infectious activity of the virus on the MARC-145 culture averaged 5.51 ± 0.45 lg TCD50/cm3 after the first three passages (after adaptation). Real-time PCR confirmed the presence of the virus genome in the test samples. Detection of the virus antigen in an infected cell culture was determined by the manifestation of a specific glow in the indirect immunofluorescence (RNIF) reaction in the cell culture monolayer fixed with acetone. It was found that FITZ-conjugate of rabbit polyclonal antibodies to pig immunoglobulins detected antigen-containing cells due to intense illumination.
ЖУРНАЛ ДЛЯ ПРОФЕССИОНАЛОВ ОТ ПРОФЕССИОНАЛОВ sensitivity of the associated antigen in detecting specific antibodies in patients with tuberculosis, as well as in animals with mixed infection, was observed, as evidenced by the study of 105 samples of animal blood serum in the farm, where a double infection was confirmed by the results of allergic, pathological and laboratory studies. Studies have shown the possibility of using of the associated antigen for the simultaneous detection of complement-binding brucellosis and tuberculosis antibodies in blood serum. The data obtained open the prospect of using of an antigenic combination in study of the diagnosis of mixed infections in animals, which is promising of increasing of economic efficiency and reducing the time for identifying epizootic and economic significant diseases.
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