A sandwich enzyme-linked immunosorbent assay (ELISA), using polyclonal antibody, was developed and compared with the commercial kit for detecting and estimating of BSA content in Measles-Mump-Rubella (MMR) vaccine samples in detection limit of nanogram level. The test depends on the capturing and detecting of BSA antigen by the polyclonal antibody. Initially, a detection range of 0-64 ng/ml was established, could be used for estimation of BSA content according to WHO requirement (50 ng/ml) in MMR vaccines. Comparative analysis of the test results for 85 MMR vaccine samples obtained with the commercial kit gave a sensitivity of 58.8% and a specificity of 97%. A high correlation (r = 0.94) was observed between BSA sandwich ELISA and commercial kit for BSA content in MMR samples. However, variations in values also were observed for the two assays. These variations may have been due to difference of upper limit of detection range of BSA content in commercial kit (32 ng/ml) and new sandwich ELISA (64 ng/ml) as well as the use of a different polyclonal antibody. In concerning the cutoff value for the WHO requirement and employment of standard solution of 64 ng/ml in developing assay, it would be adequate to use this test for assessing BSA content in viral vaccines same as MMR vaccines.
Background and Aims: The safe, potent and effective live vaccines against Measles, Mumps and Rubella as important childhood diseases have been available for several decades. Several factors can affect the thermal stability of lyophilized vaccine. Methods: In this study, the effect of residual moisture on thermal stability of 61 batches of MMR vaccine was investigated using an accelerated method that has been recommended by World Health Organization (WHO). Results: Our results suggest that the best thermal stability for the lyophilized MMR vaccine is in a range of 1.51 to 2.00% of residual moisture with the minimum decrease in the all three components of the vaccines. Conclusion: It is suggested that the lyophilizators of MMR vaccine production lines should be programmed in a manner that the best range of residual moisture achieves.
ABSTRACT:Since 1987, when chemopreventive testing programs began, more than 1,000 agents and agent combinations have been selected and evaluated in preclinical studies of chemopreventive activity against various types of cancers. In the present study we aimed to provide quantitative and qualitative characterization of biological and pharmacological activities of ICD-85 on MDA-MB-231 cell line (a highly invasive breast cancer cell line) in order to gain a better understanding of the cytotoxic and apoptotic effects of this compound. For this study, the MDA-MB-231 cell line was used and the effect of ICD-85 was assayed by measuring the activity of the cytosolic enzyme lactate dehydrogenase (LDH), released into the culture medium after membrane damage. Morphological alterations of cells were investigated in the control group and cells incubated with ICD-85 as cytotoxic agent. Results showed, in the test group, that cells incubated with 16 μg/mL of ICD-85 had decreased cytoplasmic branching. Some cells were had ruptured and lost the continuity of their surrounding membranes while some had shrunk. Cells incubated with higher doses (above16 μg/mL) showed similar changes towards cellular normality with more severity. Results obtained from the ICD-85 stability test reveal that the effect of ICD-85 on MDA-MB-231 cell line in culture medium is stable throughout the incubation time period (24 hours). It appears that ICD-85 at higher concentrations acts at the membrane level, which allows the passage of ions down the concentration gradient, resulting in osmotic changes in organelles followed by several unidentified mechanisms leading to cell death. At lower concentrations, it appears that ICD-85 can prevent cell growth by another mechanism, which may be one of the causes for apoptosis in the cell line.
Background and Aims: Adventitious agents, especially viral agents are among the most important concerns in viral vaccines. Viral contamination of biological products may arise from the original source of the reagents such as, serum, trypsin, animal or human derived media components or cell culture and working seed or cross contamination of vaccine during production. In this study, adventitious agents were evaluated in the MMR and oral polio vaccines. Methods: For detection of adventitious agents, two techniques were used. The suspensions that were harvested after inoculation on MRC-5 were neutralized with specific antisera and inoculated into Vero, HeLa and MRC-5 cells (in vitro tests). The suspensions that harvested after inoculation of CHEF primary cell culture were neutralized with specific antisera and inoculated in to Vero, MRC-5, CHEF, chorioallantoic membrane and yolk sac of SPF embryonated eggs (in vivo tests). Results: As indications of viral contamination, CPE in cell culture and the viability of egg embryo were observed and haemadsorption and haemagglutination test were performed. Finally, data were analyzed by exact binomial test (version 2.8.1). There was not any CPE in cell culture, the inoculated embryos were viable and there was not any haemadsorption & haemagglutination in all of the samples. Conclusion:The major focus of this Study will be to ensure that vaccines are devoid of adventitious agents. There were not any signs of viral agents in the samples and the preparations could be used for vaccine production.
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